Polyacid glycopeptide derivatives

ABSTRACT

Disclosed are derivatives of glycopeptides that are substituted at the C-terminus with a substituent that comprises two or more (e.g. 2, 3, 4, or 5) carboxy (CO 2 H) groups; and pharmaceutical compositions containing such glycopeptide derivatives. The disclosed glycopeptide derivatives are useful as antibacterial agents.

PRIORITY OF INVENTION

[0001] This application claims priority to U. S. Provisional ApplicationNo. 60/201,178, filed May 02, 2000; and to U. S. Provisional ApplicationNo. 60/213,415, filed Jun. 22, 2000, which applications are incorporatedherein by reference in their entirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] This invention is directed to novel polyacid derivatives ofglycopeptide antibiotics and related compounds. This invention is alsodirected to pharmaceutical compositions containing such glycopeptidederivatives, methods of using such glycopeptide derivatives asantibacterial agents, and processes and intermediates useful forpreparing such glycopeptide derivatives.

[0004] 2. Background

[0005] Glycopeptides (e.g., dalbaheptides) are a well-known class ofantibiotics produced by various microorganisms (see GlycopeptideAntibiotics, edited by R. Nagarajan, Marcel Dekker, Inc. New York(1994)). These complex multi-ring peptide compounds are very effectiveantibacterial agents against a majority of Gram-positive bacteria.Although potent antibacterial agents, the glycopeptides antibiotics arenot used in the treatment of bacterial diseases as often as otherclasses of antibiotics, such as the semi-synthetic penicillins,cephalosporins and lincomycins, due to concerns regarding toxicity.

[0006] In recent years, however, bacterial resistance to many of thecommonly-used antibiotics has developed (see J. E. Geraci et al., MayoClin. Proc. 1983, 58, 88-91; and M. Foldes, J. Antimicrob. Chemother.1983, 11, 21-26). Since glycopeptide antibiotics are often effectiveagainst these resistant strains of bacteria, glycopeptides such asvancomycin have become the drugs of last resort for treating infectionscaused by these organisms. Recently, however, resistance to vancomycinhas appeared in various microorganisms, such as vancomycin-resistantenterococci (VRE), leading to increasing concerns about the ability toeffectively treat bacterial infections in the future (see HospitalInfection Control Practices Advisory Committee, Infection ControlHospital Epidemiology, 1995, 17, 364-369; A. P. Johnson et al., ClinicalMicrobiology Rev., 1990, 3, 280-291; G. M. Eliopoulos, European J.Clinical Microbiol., Infection Disease, 1993, 12, 409-412; and P.Courvalin, Antimicrob. Agents Chemother, 1990, 34, 2291-2296).

[0007] A number of derivatives of vancomycin and other glycopeptides areknown in the art. For example, see U.S. Pat. Nos. 4,639,433; 4,643,987;4,497,802; 4,698,327; 5,591,714; 5,840,684; and 5,843,889. Otherderivatives are disclosed in EP 0 802 199; EP 0 801 075; EP 0 667 353;WO 97/28812; WO 97/38702; WO 98/52589; WO 98/52592; and in J. Amer.Chem. Soc., 1996, 118, 13107-13108; J. Amer. Chem. Soc., 1997, 119,12041-12047; and J. Amer. Chem. Soc., 1994, 116, 4573-4590.

[0008] Despite the above referenced disclosures, a need currently existsfor novel glycopeptide derivatives having effective antibacterialactivity and an improved mammalian safety profile. In particular, a needexists for glycopeptide derivatives which are effective against a widespectrum of pathogenic microorganisms, including vancomycin-resistantmicroorganisms.

SUMMARY OF THE INVENTION

[0009] The present invention provides novel polyacid glycopeptidederivatives having highly effective antibacterial activity.

[0010] Accordingly, the invention provides a compound of the invention,which is a glycopeptide substituted at the C-terminus with a substituentthat comprises two or more (e.g. 2, 3, 4, or 5) carboxy (CO₂H) groups;or a pharmaceutically acceptable salt, or stereoisomer, or prodrugthereof.

[0011] Preferably, the substituent is attached to a carbonyl group ofthe C-terminus to form an amide bond, an ester bond, or a thioesterbond. More preferably, the substituent is attached to a carbonyl groupof the C-terminus to form an amide bond. More preferably, thesubstituent comprises two carboxy (CO₂H) groups. Preferred substituentsat the C-terminus include a nitrogen-linked aspartic acid or anitrogen-linked glutamic acid.

[0012] Certain glycopeptide derivatives were disclosed by A. Malabarba,et al., J. Med. Chem., 1989, 32, 2450-2460. Accordingly, the compoundsof the invention may preferably exclude the glycopeptides 1) teicoplaninA2 substituted at the C-terminus with a nitrogen-linked glutamic acid,and 2) teicoplanin aglycon (TD) substituted at the C-terminus with anitrogen-linked glutamic acid.

[0013] Other glycopeptide derivatives are also described in U.S. patentapplication Ser. No. 09/470,209, filed Dec. 22, 1999. Accordingly, thecompounds of the invention may also preferably exclude glycopeptides offormula II:

[0014] a) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ ishydrogen;

[0015] b) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(9-hydroxydecylamino)ethyl; and R²¹is hydrogen;

[0016] c) wherein R¹⁷ is 1,4-dicarboxybutyl; R¹⁸ is hydrogen; R¹⁹ ishydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ is hydrogen; or

[0017] d) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ is—CH₂—N-(D-glucamine).

[0018] The invention also preferably exclude glycopeptides of formulaII:

[0019] e) wherein R¹⁷ is nitrogen-linked aspartic acid; R¹⁸ is hydrogen;R¹⁹ is hydrogen; R²⁰ is 2-[4-(4-chlorobenzyloxy)benzylamino]ethyl; andR²¹ is hydrogen;

[0020] f) wherein NR¹⁷ is5-(2-carboxypyrrolidin-1-ylcarbonyl)-5-(2-carboxy-3-phenylpropylamino)pentylamino;R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ ishydrogen;

[0021] g) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ is—CH₂—N—(N—CH₃-D-glucamine);

[0022] h) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ isN-[2-(2-hydroxyethoxy)ethyl]-aminomethyl; or

[0023] i) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(4-isobutylbenzyl)ethyl; and R²¹ isN-[2-(2-hydroxyethoxy)ethyl]aminomethyl.

[0024] A preferred compound of the invention is a glycopeptide offormula I:

[0025] wherein:

[0026] R¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic and —R^(a)—Y—R^(b)—(Z)_(x);or R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x);

[0027] R² is hydrogen or a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x);

[0028] R³ is a nitrogen-linked, oxygen-linked, or sulfur-linkedsubstituent comprising two or more carboxy groups;

[0029] R⁴ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, —R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide groupoptionally substituted with —R^(a)—Y—R^(b)(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x);

[0030] R⁵ is selected from the group consisting of hydrogen, halo,—CH(R^(c))—NR^(c)R^(c), —CH(R^(c))—NR^(c)—R^(e), —CH(R^(c))—R^(x),—CH(R^(c))—NR^(c)—Ra—C(═O)—R^(x), and—CH(R^(c))—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x);

[0031] R⁶ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, —R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide groupoptionally substituted with —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), or R⁵ and R⁶can be joined, together with the atoms to which they are attached, forma heterocyclic ring optionally substituted with—NR^(c)—R^(a)—Y—R^(b—(Z)) _(x);

[0032] R⁷ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, —R^(a)—Y—R^(b)—(Z)_(x), and —C(O)R^(d);

[0033] R⁸ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

[0034] R⁹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic;

[0035] R¹⁰ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic; or R⁸ and R¹⁰ arejoined to form —Ar¹—O—Ar²—, where Ar¹ and Ar² are independently aryleneor heteroarylene;

[0036] R¹¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic, or R¹⁰ and R¹¹ arejoined, together with the carbon and nitrogen atoms to which they areattached, to form a heterocyclic ring;

[0037] R¹² is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic, —C(O)R^(d), —C(NH)R^(d),—C(O)NR^(c)R^(c), —C(O)OR^(d), —C(NH)NR^(c)R^(c)and—R^(a)—Y—R^(b)—(Z)_(x), or R¹¹ and R¹² are joined, together with thenitrogen atom to which they are attached, to form a heterocyclic ring;

[0038] R¹³ is selected from the group consisting of hydrogen or —OR¹⁴;

[0039] R¹⁴ is selected from hydrogen, —C(O)R^(d) and a saccharide group;

[0040] each R^(a) is independently selected from the group consisting ofalkylene, substituted alkylene, alkenylene, substituted alkenylene,alkynylene and substituted alkynylene;

[0041] each R^(b) is independently selected from the group consisting ofa covalent bond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen;

[0042] each R^(c) is independently selected from the group consisting ofhydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclicand —C(O)R^(d);

[0043] each R^(d) is independently selected from the group consisting ofalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic;

[0044] R^(e) is a saccharide group;

[0045] each R^(f) is independently alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, or heterocyclic;

[0046] R^(x) is an N-linked amino saccharide or an N-linkedheterocyclic;

[0047] X¹, X² and X³ are independently selected from hydrogen or chloro;

[0048] each Y is independently selected from the group consisting ofoxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—, —NR^(c)C(O)—, —OSO₂—,—OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—, —SO₂NR^(c)—, —SO₂O—,—P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—, —OP(O)(OR^(c))O—,—OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—, —NR^(c)C(O)NR^(c)—,—OC(O)NR^(c)—, —C(═O)— and —NR^(c)SO₂NR^(c)—;

[0049] each Z is independently selected from hydrogen, aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic;

[0050] n is 0, 1 or 2; and

[0051] x is 1 or 2;

[0052] or a pharmaceutically acceptable salt, or stereoisomer, orprodrug thereof.

[0053] Preferably, R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), —C(O)—R^(a)—Y—R^(b)—(Z). Morepreferably R¹is an amino saccharide group substituted on the saccharidenitrogen with —CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃;—CH₂CH₂CH₂—S—(CH₂)₉CH₃; —CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH)₂)₄CH₃(trans);—CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃; —CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph;—CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph;—CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph; —CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph; or—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph. Preferably R¹ is also an aminosaccharide group substituted on the saccharide nitrogen with a4-(4-chlorophenyl)benzyl group or with a 4-(4-chlorobenzyloxy)benzylgroup.

[0054] R¹ can also be a saccharide group of the formula:

[0055] wherein R¹⁵ is —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); and R¹⁶ is hydrogen or methyl.

[0056] Preferably, R² is hydrogen.

[0057] Preferably, the substituent R³ is attached to the C-terminus toform an amide bond, an ester bond, or a thioester bond. More preferably,R³ is attached to the C-terminus to form an amide bond. More preferably,the substituent R³ comprises two carboxy (CO₂H) groups. Preferred R³substituents include a nitrogen-linked aspartic acid or a nitrogenlinked glutamic acid.

[0058] Preferably, R³ can also be a nitrogen-linked radical of formulaIII:

[0059] wherein R^(g) is a saccharide group. More preferably, R^(g) isN—(N-methyl-D-glucamine) or N-(D-glucosamine).

[0060] Preferably, R⁴, R⁶ and R⁷ are each independently selected fromhydrogen or —C(O)R^(d). More preferably, R⁴, R⁶ and R⁷ are eachhydrogen.

[0061] Preferably R⁵ is hydrogen, —CH₂—NHR^(c), —CH₂—NR^(c)R^(e) or—CH₂—NH—R^(a)—Y—R^(b)—(Z)_(x). R⁵ can also preferably be hydrogen;—CH₂—N—(N—CH₃-D-glucamine); —CH₂—NH—CH₂CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—NHC(O)—(CH₂)₃COOH; —CH₂—NH—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—COOH; —CH₂—NH—(CH₂)₅COOH; —CH₂-(morpholin-4-yl);—CH₂—NH—CH₂CH₂—O—CH₂CH₂OH; —CH₂—NH—CH₂CH(OH)—CH₂OH; —CH₂—N[CH₂CH₂OH]₂;—CH₂—NH—(CH₂)₃—N(CH₃)₂; —CH₂—N[(CH₂)₃—N(CH₃)₂]₂;—CH₂—NH—(CH₂)₃-(imidazol-1-yl); —CH₂—NH—(CH₂)₃-(morpholin-4-yl);—CH₂—NH—(CH₂)₄—NHC(NH)NH₂; —CH₂—N—(2-amino-2-deoxygluconic acid);—CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; —CH₂—NH—CH(COOH)CH₂COOH;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₈CH₃;—CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₉CH₃; —CH₂—NH—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃;—CH₂—NH—CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂—NH—CH₂CH₂—O—CH₂CH₂OH;—CH₂—NH—CH₂CH₂C(O)—N-(D-glucosamine);—CH₂—NH-(6-oxo-[1,3]oxazinan-3-yl); —CH₂—NH—CH₂CH₂—S—(CH₂)₇CH₃;—CH₂—NH—CH₂CH₂—S—(CH₂)₈CH₃; —CH₂—NH—CH₂CH₂—S—(CH₂)₉CH₃;—CH₂—NH—CH₂CH₂—S—(CH₂)₁₁CH₃; —CH₂—NH—CH₂CH₂—S—(CH₂)₆Ph;—CH₂—NH—CH₂CH₂—S—(CH₂)₈Ph; —CH₂—NH—CH₂CH₂—S—(CH₂)₁₀Ph;—CH₂—NH—CH₂CH₂—S—CH₂-(4-(4-CF₃-Ph)Ph); —CH₂—NH—CH₂CH₂—NH—(CH₂)₁₁CH₃; or—CH₂—NH—(CH₂)₅—COOH;.

[0062] Preferably, R⁸ is —CH₂C(O)NH₂, —CH₂COOH, benzyl, 4-hydroxyphenylor 3-chloro-4-hydroxyphenyl.

[0063] Preferably, R⁹ is hydrogen or alkyl.

[0064] Preferably, R¹⁰ is alkyl or substituted alkyl. More preferably,R¹⁰ is the side-chain of a naturally occurring amino acid, such asisobutyl.

[0065] Preferably, R¹¹ is hydrogen or alkyl.

[0066] Preferably, R¹² is hydrogen, alkyl, substituted alkyl or—C(O)R^(d). R¹² can also preferably be hydrogen; —CH₂COOH;—CH₂—[CH(OH)]₅CH₂OH; —CH₂CH(OH)CH₂OH; —CH₂CH₂NH₂; —CH₂C(O)OCH₂CH₃;—CH₂-(2-pyridyl); —CH₂—[CH(OH)]₄COOH; —CH₂-(3-carboxyphenyl);(R)—C(O)CH(NH₂)(CH₂)₄NH₂; —C(O)Ph; —C(O)CH₂NHC(O)CH₃;E-CH₂CH₂—S—(CH₂)₃CH═CH(CH₂)₄CH₃; or —C(O)CH₃.

[0067] Preferably, X¹ and X² are each chloro.

[0068] Preferably, X³ is hydrogen.

[0069] Preferably each Y is independently selected from the groupconsisting of oxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—,—NR^(c)C(O)—, —OSO₂—, —OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—,—SO₂NR^(c)—, —SO₂O—, —P(O)(OR^(c))O—, —P(O)(OR^(c))NR^(c)—,—OP(O)(OR^(c))O—, —OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—,—NR^(c)C(O)NR^(c)—, —OC(O)NR^(c)— and —NR^(c)SO₂NR^(c)—;

[0070] Preferably, n is 0 or 1, and more preferably, n is 1.

[0071] Another preferred compound of the invention is a glycopeptide offormula II:

[0072] wherein

[0073] R¹⁷ is a dicarboxy-substituted alkyl group having from 3 to 10carbon atoms;

[0074] R¹⁸ is selected from the group consisting of hydrogen and alkyl;

[0075] R¹⁹ is hydrogen;

[0076] R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x);

[0077] R²¹ is hydrogen;

[0078] R^(a) is selected from the group consisting of alkylene,substituted alkylene, alkenylene, substituted alkenylene, alkynylene andsubstituted alkynylene;

[0079] R^(b) is selected from the group consisting of a covalent bond,alkylene, substituted alkylene, alkenylene, substituted alkenylene,alkynylene and substituted alkynylene, provided R^(b) is not a covalentbond when Z is hydrogen;

[0080] Y is selected from the group consisting of sulfur, —S(O)— and—SO₂—;

[0081] each Z is independently selected from hydrogen, aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic; and

[0082] x is 1 or 2;

[0083] or a pharmaceutically acceptable salt, or stereoisomer, orprodrug thereof.

[0084] For a compound of formula II, R^(a) is preferably selected fromalkylene having from 1 to 10 carbon atoms. For example, R^(a) can beethylene (—CH₂CH₂—), propylene (—CH₂CH₂CH₂—) or butylene(—CH₂CH₂CH₂CH₂—).

[0085] For a compound of formula II, Z is preferably hydrogen and R^(b)is alkylene of from 8 to 12 carbon atoms. More preferably, R^(b) can bea covalent bond, methylene, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉— or—(CH₂)₁₀—. Additionally, R^(b) and Z can preferably form an n-octyl,n-nonyl, n-decyl, n-undecyl or n-dodecyl group.

[0086] For a compound of formula II, Z is preferably aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic, and R^(b) is a covalent bondor alkylene of from 1 to 10 carbon atoms. More preferably, Z can bearyl. Preferred aryl groups include phenyl, substituted phenyl,biphenyl, substituted biphenyl and terphenyl groups. Most preferably, Zcan be phenyl, 4-isobutylphenyl, 4′-chlorobiphenyl-4-yl,4′-trifluoromethylbiphenyl-4-yl, 4-(naphth-2-yl)phenyl,4-(2-phenylethynyl)phenyl, 4-(3,4-dichlorobenzyloxy)-phenyl, orp-terphenyl.

[0087] For a compound of formula II, x is preferably 1.

[0088] For a compound of formula II, Y is preferably sulfur.

[0089] For a compound of formula I or II, —R^(a)—Y—R^(b) —(Z)_(x) ispreferably selected from the group consisting of: —CH₂CH₂—S—(CH₂)₈CH₃;—CH₂CH₂—S—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃;—CH₂CH₂CH₂—S—(CH₂)₉CH₃; —CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃ (trans);—CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃; —CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph;—CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph; and—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph. More preferably,—R^(a)—Y—R^(b)(Z)_(x) is —CH₂CH₂—S—(CH₂)₉CH₃.

[0090] For a compound of formula II, R¹⁷ is preferably adicarboxy-substituted alkyl group having from 4 to 6 carbon atoms. Morepreferably R¹⁷ is —CH₂(COOH)CH₂COOH or —CH₂(COOH)CH₂CH₂COOH. Mostpreferably, R¹⁷ is —CH₂(COOH)CH₂COOH.

[0091] For a compound of formula II, R¹⁸ is preferably hydrogen.

[0092] A preferred compound of formula II is a compound wherein—R^(a)—Y—R^(b)—(Z)_(x) is —CH₂CH₂—S—(CH₂)₉CH₃; R¹⁷ is —CH₂(COOH)CH₂COOH;and R¹⁸ is hydrogen.

[0093] A preferred value for R¹⁵, R²⁰, or —R^(a)—Y—R^(b)—(Z)_(x) is—CH₂CH₂—NH—(CH₂)₉CH₃; —CH₂CH₂CH₂—NH—(CH₂)₈CH₃;—CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃; —CH₂CH₂—NHSO₂—(CH₂)₉CH₃;—CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃; —CH₂CH₂CH₂—S—(CH₂)₉CH₃;—CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃(trans); —CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃;—CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph; —CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph; —CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph;—CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph; or—CH₂CH₂CH₂-NHSO₂-4-(naphth-2yl)-Ph. Another preferred value for R¹⁵ orR²⁰ is 4-(4-chlorophenyl)benzyl or 4-(4-chlorobenzyloxy)benzyl.

[0094] The invention also provides a pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and a therapeuticallyeffective amount of a compound of the invention. In one preferredembodiment, the pharmaceutically acceptable carrier comprises an aqueouscyclodextrin solution. Preferably, the cyclodextrin ishydroxypropyl-β-cyclodextrin or sulfobutyl ether β-cyclodextrin. Morepreferably, the cyclodextrin is hydroxypropyl-β-cyclodextrin.

[0095] The compounds of the invention are highly effective antibacterialagents. Accordingly, the invention also provides a method of treating amammal having a bacterial disease, comprising administering to themammal a therapeutically effective amount of a compound of theinvention. The invention also provides a method of treating a mammalhaving a bacterial disease, comprising administering to the mammal atherapeutically effective amount of a pharmaceutical composition of theinvention.

[0096] The invention also provides processes and intermediates usefulfor preparing compounds of the invention, which processes andintermediates are described further herein.

[0097] The invention also provides a compound of the invention asdescribed herein for use in medical therapy, as well as the use of acompound of the invention in the manufacture of a formulation ormedicament for treating a bacterial disease in a mammal.

[0098] Preferred compounds of the invention are the compounds of formulaII shown in Table I below wherein R¹⁹ and R²¹ are each hydrogen. TABLE IPreferred Compounds of Formula II Com- pound NR¹⁷R¹⁸ R²⁰ 1 N-linkedaspartic acid CH₃(CH₂)₉— 2 N-linked aspartic acid CH₃(CH₂)₁₀— 3 N-linkedaspartic acid CH₃(CH₂)₁₁— 4 N-linked aspartic acid CH₃(CH₂)₁₂— 5N-linked aspartic acid CH₃(CH₂)₉SCH₂CH₂— 6 Formula IV, wherein R^(g)CH₃(CH₂)₉SCH₂CH₂— is N-(D-glucosamine). 7 N-linked aspartic acidCH₃(CH₂)₉OCH₂CH₂— 8 N-linked aspartic acid 4-(4-chlorophenyl)benzyl 9N-linked aspartic acid 2-(4-(4-chlorophenyl)benzylthio)ethyl 10 N-linkedaspartic acid 2-(4-(4-chlorophenyl)benzyloxy)ethyl 11 N-linked asparticacid 4-(4′-chlorobiphenyl)butyl 12 N-linked glutamic acid4-(4-chlorophenyl)benzyl 13 N-linked glutamic acid CH₃(CH₂)₉NHCH₂CH₂— 14Formula III, wherein R^(g) CH₃(CH₂)₉NHCH₂CH₂— is N-(D-glucamine) 15Formula III, wherein R^(g) CH₃(CH₂)₉OCH₂CH₂— is N-(D-glucamine)

[0099] Another preferred group of compounds of the invention areC-terminus polyacid derivatives of the glycopeptide antibiotic A82846B(also known as chloroorienticin A oy LY264826). See for example R.Nagarajan et al., J. Org. Chem., 1988, 54, 983-986; and N. Tsuji et al.,J. Antibiot., 1988, 41, 819-822. The structure of this glycopeptide issimilar to vancomycin, except A82846B contains an additional amino sugar(i.e. 4-epi-vancosamine attached at the R² position in formula I.) andfurther contains 4-epi-vancosamine in place of vancosamine in thedisaccharide moiety attached at the R¹ position in formula I. Forexample, a preferred group of compounds are N-alkylated derivatives ofA82846B having a polyacid group at the C-terminus. N-alkylatedderivatives of A82846B are known in the art and are described, forexample, in U.S. Pat. No. 5,840,684. C-Terminus polyacid derivatives ofthese compounds can readily be prepared using the procedures describedherein.

[0100] A particularly preferred group of compounds of the invention areA82846B derivatives having a 4-(4-chlorophenyl)benzyl group or a4-(4-chlorobenzyloxy)benzyl group attached at the amino group of the4-epi-vancosamine of the disaccharide moiety and having a groupsubstituted at the C-terminus that comprises two or more carboxy group.Preferred polycarboxy substituents for A82846B derivatives include anitrogen-linked aspartic acid, a nitrogen linked glutamic acid, and anitrogen-linked radical of formula III as described herein.

DETAILED DESCRIPTION OF THE INVENTION

[0101] This invention relates to novel derivatives of glycopeptideantibiotics comprising a polyacid containing group at the C-terminus(compounds of the invention), as well as to compositions comprising suchcompounds and to therapeutic methods comprising the administration ofsuch compounds. When describing the compounds, compositions and methodsof the invention, the following terms have the following meanings,unless otherwise indicated.

[0102] Definitions

[0103] The term “alkyl” refers to a monoradical branched or unbranchedsaturated hydrocarbon chain preferably having from 1 to 40 carbon atoms,more preferably 1 to 10 carbon atoms, and even more preferably 1 to 6carbon atoms. This term is exemplified by groups such as methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, n-hexyl, n-decyl, tetradecyl,and the like.

[0104] The term “substituted alkyl” refers to an alkyl group as definedabove, having from 1 to 8 substituents, preferably 1 to 5 substituents,and more preferably 1 to 3 substituents, selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, guanido, halogen, hydroxyl, keto, thioketo,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₃H, and —SO₂-heteroaryl.

[0105] The term “alkylene” refers to a diradical of a branched orunbranched saturated hydrocarbon chain, preferably having from 1 to 40carbon atoms, preferably 1-10 carbon atoms, more preferably 1-6 carbonatoms. This term is exemplified by groups such as methylene (—CH₂—),ethylene (—CH₂CH₂—), the propylene isomers (e.g., —CH₂CH₂CH₂— and—CH(CH₃)CH₂—) and the like.

[0106] The term “substituted alkylene” refers to an alkylene group, asdefined above, having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkylene groupsinclude those where 2 substituents on the alkylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkylene group. Preferably such fused groups contain from 1 to 3fused ring structures. Additionally, the term substituted alkyleneincludes alkylene groups in which from 1 to 5 of the alkylene carbonatoms are replaced with oxygen, sulfur or —NR— where R is hydrogen oralkyl. Examples of substituted alkylenes are chloromethylene (—CH(Cl)—),aminoethylene (—CH(NH₂)CH2—), 2-carboxypropylene isomers(—CH₂CH(CO₂H)CH₂—), ethoxyethyl (—CH₂CH₂O—CH₂CH₂—) and the like.

[0107] The term “alkaryl” refers to the groups -alkylene-aryl and-substituted alkylene-aryl where alkylene, substituted alkylene and arylare defined herein. Such alkaryl groups are exemplified by benzyl,phenethyl and the like.

[0108] The term “alkoxy” refers to the groups alkyl-O—, alkenyl-O—,cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, where alkyl, alkenyl,cycloalkyl, cycloalkenyl, and alkynyl are as defined herein. Preferredalkoxy groups are alkyl-O— and include, by way of example, methoxy,ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0109] The term “substituted alkoxy” refers to the groups substitutedalkyl-O—, substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

[0110] The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylalkoxy groups are alkylene-O-alkyl and include, by way ofexample, methylenemethoxy (—CH₂OCH₃), ethylenemethoxy (—CH₂CH₂OCH₃),n-propylene-iso-propoxy (—CH₂CH₂CH₂OCH(CH₃)₂), methylene-t-butoxy(—CH₂—O—C(CH₃)₃) and the like.

[0111] The term “alkylthioalkoxy” refers to the group -alkylene-S-alkyl,alkylene-S-substituted alkyl, substituted alkylene-S-alkyl andsubstituted alkylene-S-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.Preferred alkylthioalkoxy groups are alkylene-S-alkyl and include, byway of example, methylenethiomethoxy (—CH₂SCH₃), ethylenethiomethoxy(—CH₂CH₂SCH₃), n-propylene-iso-thiopropoxy (—CH₂CH₂CH₂ SCH(CH₃)₂),methylene-t-thiobutoxy (—CH₂SC(CH₃)₃) and the like.

[0112] The term “alkenyl” refers to a monoradical of a branched orunbranched unsaturated hydrocarbon group preferably having from 2 to 40carbon atoms, more preferably 2 to 10 carbon atoms and even morepreferably 2 to 6 carbon atoms and having at least 1 and preferably from1-6 sites of vinyl unsaturation. Preferred alkenyl groups includeethenyl (—CH═CH₂), n-propenyl (—CH₂CH═CH₂), iso-propenyl (—C(CH₃)═CH₂),and the like.

[0113] The term “substituted alkenyl” refers to an alkenyl group asdefined above having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

[0114] The term “alkenylene” refers to a diradical of a branched orunbranched unsaturated hydrocarbon group preferably having from 2 to 40carbon atoms, more preferably 2 to 10 carbon atoms and even morepreferably 2 to 6 carbon atoms and having at least 1 and preferably from1-6 sites of vinyl unsaturation. This term is exemplified by groups suchas ethenylene (—CH═CH—), the propenylene isomers (e.g., —CH₂CH═CH— and—C(CH₃)═CH—) and the like.

[0115] The term “substituted alkenylene” refers to an alkenylene groupas defined above having from 1 to 5 substituents, and preferably from 1to 3 substituents, selected from the group consisting of alkoxy,substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substitutedamino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen,hydroxyl, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl. Additionally, such substituted alkenylene groupsinclude those where 2 substituents on the alkenylene group are fused toform one or more cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heterocyclic or heteroaryl groups fusedto the alkenylene group.

[0116] The term “alkynyl” refers to a monoradical of an unsaturatedhydrocarbon preferably having from 2 to 40 carbon atoms, more preferably2 to 20 carbon atoms and even more preferably 2 to 6 carbon atoms andhaving at least 1 and preferably from 1-6 sites of acetylene (triplebond) unsaturation. Preferred alkynyl groups include ethynyl (—C≡CH),propargyl (—CH₂C≡CH) and the like.

[0117] The term “substituted alkynyl” refers to an alkynyl group asdefined above having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

[0118] The term “alkynylene” refers to a diradical of an unsaturatedhydrocarbon preferably having from 2 to 40 carbon atoms, more preferably2 to 10 carbon atoms and even more preferably 2 to 6 carbon atoms andhaving at least 1 and preferably from 1-6 sites of acetylene (triplebond) unsaturation. Preferred alkynylene groups include ethynylene(—C≡C—), propargylene (—CH₂C≡C—) and the like.

[0119] The term “substituted alkynylene” refers to an alkynylene groupas defined above having from 1 to 5 substituents, and preferably 1 to 3substituents, selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl The term “acyl” refers to the groups HC(O)—,alkyl-C(O)—, substituted alkyl-C(O)—, cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—,aryl-C(O)—, heteroaryl-C(O)— and heterocyclic-C(O)— where alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic are asdefined herein.

[0120] The term “acylamino” or “aminocarbonyl” refers to the group—C(O)NRR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, heterocyclic or where both R groups are joinedto form a heterocyclic group (e.g., morpholino) wherein alkyl,substituted alkyl, aryl, heteroaryl and heterocyclic are as definedherein.

[0121] The term “aminoacyl” refers to the group —NRC(O)R where each R isindependently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, orheterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

[0122] The term “aminoacyloxy” or “alkoxycarbonylamino” refers to thegroup —NRC(O)OR where each R is independently hydrogen, alkyl,substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl,substituted alkyl, aryl, heteroaryl and heterocyclic are as definedherein.

[0123] The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclic-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclic are as defined herein.

[0124] The term “aryl” refers to an unsaturated aromatic carbocyclicgroup of from 6 to 20 carbon atoms having a single ring (e.g., phenyl)or multiple condensed (fused) rings, wherein at least one ring isaromatic (e.g., naphthyl, dihydrophenanthrenyl, fluorenyl, or anthryl).Preferred aryls include phenyl, naphthyl and the like.

[0125] Unless otherwise constrained by the definition for the arylsubstituent, such aryl groups can optionally be substituted with from 1to 5 substituents, preferably 1 to 3 substituents, selected from thegroup consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, sulfonamide,thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy,—SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy.

[0126] The term “aryloxy” refers to the group aryl-O— wherein the arylgroup is as defined above including optionally substituted aryl groupsas also defined above.

[0127] The term “arylene” refers to the diradical derived from aryl(including substituted aryl) as defined above and is exemplified by1,2-phenylene, 1,3-phenylene, 1,4-phenylene, 1,2-naphthylene and thelike.

[0128] The term “amino” refers to the group —NH₂.

[0129] The term “substituted amino” refers to the group —NRR where eachR is independently selected from the group consisting of hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl and heterocyclic provided thatboth R's are not hydrogen.

[0130] “Amino acid” refers to any of the naturally occurring amino acids(e.g. Ala, Arg, Asn, Asp, Cys, Glu, Gln, Gly, His, Hyl, Hyp, Ile, Leu,Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val) in D, L, or DL form.The side chains of naturally occurring amino acids are well known in theart and include, for example, hydrogen (e.g., as in glycine), alkyl(e.g., as in alanine, valine, leucine, isoleucine, proline), substitutedalkyl (e.g., as in threonine, serine, methionine, cysteine, asparticacid, asparagine, glutamic acid, glutamine, arginine, and lysine),alkaryl (e.g., as in phenylalanine and tryptophan), substitutedarylalkyl (e.g., as in tyrosine), and heteroarylalkyl (e.g., as inhistidine).

[0131] The term “carboxy” refers to —COOH.

[0132] The term “C-terminus” as it relates to a glycopeptide is wellunderstood in the art. For example, for a glycopeptide of formula I, theC-terminus is the position substituted by the group R³.

[0133] The term “dicarboxy-substituted alkyl” refers to an alkyl groupsubstituted with two carboxy groups. This term includes, by way ofexample, —CH₂(COOH)—CH₂COOH and —CH₂(COOH)CH₂CH₂COOH.

[0134] The term “carboxyalkyl” or “alkoxycarbonyl” refers to the groups“—C(O)O-alkyl”, “—C(O)O-substituted alkyl”, “—C(O)O-cycloalkyl”,“—C(O)O— substituted cycloalkyl”, “—C(O)O-alkenyl”, “—C(O)O-substitutedalkenyl”, “—C(O)O-alkynyl” and “—C(O)O-substituted alkynyl” where alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, alkynyl and substituted alkynyl alkynyl are asdefined herein.

[0135] The term “cycloalkyl” refers to cyclic alkyl groups of from 3 to20 carbon atoms having a single cyclic ring or multiple condensed rings.Such cycloalkyl groups include, by way of example, single ringstructures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, andthe like, or multiple ring structures such as adamantanyl, and the like.

[0136] The term “substituted cycloalkyl” refers to cycloalkyl groupshaving from 1 to 5 substituents, and preferably 1 to 3 substituents,selected from the group consisting of alkoxy, substituted alkoxy,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

[0137] The term “cycloalkenyl” refers to cyclic alkenyl groups of from 4to 20 carbon atoms having a single cyclic ring and at least one point ofinternal unsaturation. Examples of suitable cycloalkenyl groups include,for instance, cyclobut-2-enyl, cyclopent-3-enyl, cyclooct-3-enyl and thelike.

[0138] The term “substituted cycloalkenyl” refers to cycloalkenyl groupshaving from 1 to 5 substituents, and preferably 1 to 3 substituents,selected from the group consisting of alkoxy, substituted alkoxy,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryland —SO₂-heteroaryl.

[0139] The term “halo” or “halogen” refers to fluoro, chloro, bromo andiodo.

[0140] “Haloalkyl” refers to alkyl as defined herein substituted by 1-4halo groups as defined herein, which may be the same or different.Representative haloalkyl groups include, by way of example,trifluoromethyl, 3-fluorododecyl, 12,12,12-trifluorododecyl,2-bromooctyl, 3-bromo-6-chloroheptyl, and the like.

[0141] The term “heteroaryl” refers to an aromatic group of from 1 to 15carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen andsulfur within at least one ring (if there is more than one ring).

[0142] Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 5 substituents, preferably 1 to 3 substituents, selected from thegroup consisting of acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy,alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, heteroaryloxy,heterocyclic, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.Preferred aryl substituents include alkyl, alkoxy, halo, cyano, nitro,trihalomethyl, and thioalkoxy. Such heteroaryl groups can have a singlering (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl,pyrrolyl and furyl.

[0143] “Heteroarylalkyl” refers to (heteroaryl)alkyl- where heteroaryland alkyl are as defined herein. Representative examples include2-pyridylmethyl and the like.

[0144] The term “heteroaryloxy” refers to the group heteroaryl-O—.

[0145] The term “heteroarylene” refers to the diradical group derivedfrom heteroaryl (including substituted heteroaryl), as defined above,and is exemplified by the groups 2,6-pyridylene, 2,4-pyridiylene,1,2-quinolinylene, 1,8-quinolinylene, 1,4-benzofuranylene,2,5-pyridnylene, 2,5-indolenyl and the like.

[0146] The term “heterocycle” or “heterocyclic” refers to a monoradicalsaturated or unsaturated group having a single ring or multiplecondensed rings, from 1 to 40 carbon atoms and from 1 to 10 heteroatoms, preferably 1 to 4 heteroatoms, selected from nitrogen, sulfur,phosphorus, and/or oxygen within the ring.

[0147] Unless otherwise constrained by the definition for theheterocyclic substituent, such heterocyclic groups can be optionallysubstituted with 1 to 5, and preferably 1 to 3 substituents, selectedfrom the group consisting of alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy,carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol,thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, oxo (O═), and—SO₂-heteroaryl. Such heterocyclic groups can have a single ring ormultiple condensed rings. Preferred heterocyclics include morpholino,piperidinyl, and the like.

[0148] Examples of nitrogen heterocycles and heteroaryls include, butare not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, indoline, morpholino, piperidinyl,tetrahydrofuranyl, and the like as well as N-alkoxy-nitrogen containingheterocycles.

[0149] Another class of heterocyclics is known as “crown compounds”which refers to a specific class of heterocyclic compounds having one ormore repeating units of the formula [—(CH₂—)_(a)A—] where a is equal toor greater than 2, and A at each separate occurrence can be O, N, S orP. Examples of crown compounds include, by way of example only,[—(CH₂)₃—NH—]₃, [—((CH₂)₂—O)₄—((CH₂)₂—NH)₂] and the like. Typically suchcrown compounds can have from 4 to 10 heteroatoms and 8 to 40 carbonatoms.

[0150] The term “heterocyclooxy” refers to the group heterocyclic-O—.

[0151] The term “thioheterocyclooxy” refers to the groupheterocyclic-S—.

[0152] The term “oxyacylamino” or “aminocarbonyloxy” refers to the group—OC(O)NRR where each R is independently hydrogen, alkyl, substitutedalkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substitutedalkyl, aryl, heteroaryl and heterocyclic are as defined herein.

[0153] The term “prodrug” is well understood in the art and includescompounds that are converted to pharmaceutically active compounds of theinvention in a mammalian system. For example, see Remington'sPharmaceutical Sciences, 1980, vol 16, Mack Publishing Company, Easton,Pa., 61 and 424.

[0154] The term “saccharide group” refers to an oxidized, reduced orsubstituted saccharide monoradical covalently attached to theglycopeptide or other compound via any atom of the saccharide moiety,preferably via the aglycone carbon atom. The term includesamino-containing saccharide groups. Representative saccharides include,by way of illustration, hexoses such as D-glucose, D-mannose, D-xylose,D-galactose, vancosamine, 3-desmethyl-vancosamine, 3-epi-vancosamine,4-epi-vancosamine, acosamine, actinosamine, daunosamine,3-epi-daunosamine, ristosamine, D-glucamine, N-methyl-D-glucamine,D-glucuronic acid, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine,sialyic acid, iduronic acid, L-fucose, and the like; pentoses such asD-ribose or D-arabinose; ketoses such as D-ribulose or D-fructose;disaccharides such as 2-O-(α-L-vancosaminyl)-β-D-glucopyranose,2-O-(3-desmethyl-α-L-vancosaminyl)-β-D-glucopyranose, sucrose, lactose,or maltose; derivatives such as acetals, amines, acylated, sulfated andphosphorylated sugars; oligosaccharides having from 2 to 10 saccharideunits. For the purposes of this definition, these saccharides arereferenced using conventional three letter nomenclature and thesaccharides can be either in their open or preferably in their pyranoseform.

[0155] The term “amino-containing saccharide group” or “aminosaccharide” refers to a saccharide group having an amino substituent.Representative amino-containing saccharides include L-vancosamine,3-desmethyl-vancosamine, 3-epi-vancosamine, 4-epi-vancosamine,acosamine, actinosamine, daunosamine, 3-epi-daunosamine, ristosamine,N-methyl-D-glucamine and the like.

[0156] The term “spiro-attached cycloalkyl group” refers to a cycloalkylgroup attached to another ring via one carbon atom common to both rings.

[0157] The term “stereoisomer” as it relates to a given compound is wellunderstood in the art, and refers another compound having the samemolecular formula, wherein the atoms making up the other compound differin the way they are oriented in space, but wherein the atoms in theother compound are like the atoms in the given compound with respect towhich atoms are joined to which other atoms (e.g. an enantiomer, adiastereomer, or a geometric isomer). See for example, Morrison andBoyde Organic Chemistry, 1983, 4th ed., Allyn and Bacon, Inc., Boston,Mass., page 123.

[0158] The term “sulfonamide” refers to a group of the formula —SO₂NRR,where each R is independently hydrogen, alkyl, substituted alkyl, aryl,heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

[0159] The term “thiol” refers to the group —SH.

[0160] The term “thioalkoxy” refers to the group —S-alkyl.

[0161] The term “substituted thioalkoxy” refers to the group—S-substituted alkyl.

[0162] The term “thioaryloxy” refers to the group aryl-S— wherein thearyl group is as defined above including optionally substituted arylgroups also defined above.

[0163] The term “thioheteroaryloxy” refers to the group heteroaryl-S—wherein the heteroaryl group is as defined above including optionallysubstituted aryl groups as also defined above.

[0164] The term “thioether derivatives” when used to refer to theglycopeptide compounds of this invention includes thioethers (—S—),sulfoxides (—SO—) and sulfones (—SO₂—).

[0165] As to any of the above groups which contain one or moresubstituents, it is understood, of course, that such groups do notcontain any substitution or substitution patterns which are stericallyimpractical and/or synthetically non-feasible. In addition, thecompounds of this invention include all stereochemical isomers arisingfrom the substitution of these compounds.

[0166] “Cyclodextrin” refers to cyclic molecules containing six or moreα-D-glucopyranose units linked at the 1,4 positions by a linkages as inamylose. β-Cyclodextrin or cycloheptaamylose contains sevena-D-glucopyranose units. As used herein, the term “cyclodextrin” alsoincludes cyclodextrin derivatives such as hydroxypropyl and sulfobutylether cyclodextrins. Such derivatives are described for example, in U.S.Pat. Nos. 4,727,064 and 5,376,645. One preferred cyclodextrin ishydroxypropyl β-cyclodextrin having a degree of substitution of fromabout 4.1-5.1 as measured by FTIR. Such a cyclodextrin is available fromCerestar (Hammond, Ind., USA) under the name Cavitron™ 82003.

[0167] “Glycopeptide” refers to oligopeptide (e.g. a heptapeptide)antibiotic, characterized by a multi-ring peptide core optionallysubstituted with saccharide groups, such as vancomycin. Examples ofglycopeptides included in this definition may be found in “GlycopeptidesClassification, Occurrence, and Discovery”, by Raymond C. Rao and LouiseW. Crandall, (“Drugs and the Pharmaceutical Sciences” Volume 63, editedby Ramakrishnan Nagarajan, published by Marcal Dekker, Inc.). Additionalexamples of glycopeptides are disclosed in U.S. Pat. Nos. 4,639,433;4,643,987; 4,497,802; 4,698,327; 5,591,714; 5,840,684; and 5,843,889; inEP 0 802 199; EP 0 801 075; EP 0 667 353; WO 97/28812; WO 97/38702; WO98/52589; WO 98/52592; and in J. Amer. Chem. Soc., 1996, 118,13107-13108; J. Amer. Chem. Soc., 1997, 119, 12041-12047; and J. Amer.Chem. Soc., 1994, 116, 4573-4590. Representative glycopeptides includethose identified as A477, A35512, A40926, A41030, A42867, A47934,A80407, A82846, A83850, A84575, AB-65, Actaplanin, Actinoidin, Ardacin,Avoparcin, Azureomycin, Balhimycin, Chloroorientiein, Chloropolysporin,Decaplanin, N-demethyl-vancomycin, Eremomycin, Galacardin, Helvecardin,Izupeptin, Kibdelin, LL-AM374, Mannopeptin, MM45289, MM47756, MM47761,MM49721, MM47766, MM55260, MM55266, MM55270, MM56597, MM56598, OA-7653,Orenticin, Parvodicin, Ristocetin, Ristomycin, Synmonicin, Teicoplanin,UK-68597, UK-69542, UK-72051, Vancomycin, and the like. The term“glycopeptide” as used herein is also intended to include the generalclass of peptides disclosed above on which the sugar moiety is absent,i.e. the aglycone series of glycopeptides. For example, removal of thedisaccharide moiety appended to the phenol on vancomycin by mildhydrolysis gives vancomycin aglycone. Also within the scope of theinvention are glycopeptides that have been further appended withadditional saccharide residues, especially aminoglycosides, in a mannersimilar to vancosamine.

[0168] “Vancomycin” refers to a glycopeptide antibiotic having theformula:

[0169] When describing vancomycin derivatives, the term “N^(van)—”indicates that a substituent is covalently attached to the amino groupof the vacosamine moiety of vacomycin. Similarly, the term “N^(leu)—”indicates that a substituent is covalently attached to the amino groupof the leucine moiety of vancomycin.

[0170] “Optional” or “optionally” means that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where said event or circumstance occurs and instancesin which it does not. For example, “optionally substituted” means that agroup may or may not be substituted with the described substituent.

[0171] As used herein, the terms “inert organic solvent” or “inertsolvent” or “inert diluent” mean a solvent or diluent which isessentially inert under the conditions of the reaction in which it isemployed as a solvent or diluent. Representative examples of materialswhich may be used as inert solvents or diluents include, by way ofillustration, benzene, toluene, acetonitrile, tetrahydrofuran (“THF”),dimethylformamide (“DMF”), chloroform (“CHCl₃”), methylene chloride (ordichloromethane or ″CH₂Cl₂), diethyl ether, ethyl acetate, acetone,methylethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

[0172] The term “nitrogen-linked” or “N-linked” means a group orsubstituent is attached to the remainder of a compound (e.g. a compoundof formula I) through a bond to a nitrogen of the group or substituent.The term “oxygen-linked” means a group or substituent is attached to theremainder of a compound (e.g. a compound of formula I) through a bond toan oxygen of the group or substituent. The term “sulfur-linked” means agroup or substituent is attached to the remainder of a compound (e.g. acompound of formula I) through a bond to a sulfur of the group orsubstituent.

[0173] “Pharmaceutically acceptable salt” means those salts which retainthe biological effectiveness and properties of the parent compounds andwhich are not biologically or otherwise harmful as the dosageadministered. The compounds of this invention are capable of formingboth acid and base salts by virtue of the presence of amino and carboxygroups respectively.

[0174] Pharmaceutically acceptable base addition salts may be preparedfrom inorganic and organic bases. Salts derived from inorganic basesinclude, but are not limited to, the sodium, potassium, lithium,ammonium, calcium, and magnesium salts. Salts derived from organic basesinclude, but are not limited to, salts of primary, secondary andtertiary amines, substituted amines including naturally-occurringsubstituted amines, and cyclic amines, including isopropylamine,trimethyl amine, diethylamine, triethylamine, tripropylamine,ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine,histidine, caffeine, procaine, hydrabamine, choline, betaine,ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines,piperazine, piperidine, and N-ethylpiperidine. It should also beunderstood that other carboxylic acid derivatives would be useful in thepractice of this invention, for example carboxylic acid amides,including carboxamides, lower alkyl carboxamides, di(lower alkyl)carboxamides, and the like.

[0175] Pharmaceutically acceptable acid addition salts may be preparedfrom inorganic and organic acids. Salts derived from inorganic acidsinclude hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid and the like.

[0176] The compounds of this invention typically contain one or morechiral centers. Accordingly, this invention is intended to includeracemic mixtures, diasteromers, enantiomers and mixture enriched in oneor more steroisomer. The scope of the invention as described and claimedencompasses the racemic forms of the compounds as well as the individualenantiomers and non-racemic mixtures thereof.

[0177] The term “treatment” as used herein includes any treatment of acondition or disease in an animal, particularly a mammal, moreparticularly a human, and includes:

[0178] (i) preventing the disease or condition from occurring in asubject which may be predisposed to the disease but has not yet beendiagnosed as having it;

[0179] (ii) inhibiting the disease or condition, i.e. arresting itsdevelopment; relieving the disease or condition, i.e. causing regressionof the condition; or relieving the conditions caused by the disease,i.e. symptoms of the disease.

[0180] The term “disease state which is alleviated by treatment with abroad spectrum antibacterial” or “bacterial disease” as used herein isintended to cover all disease states which are generally acknowledged inthe art to be usefully treated with a broad spectrum antibacterial ingeneral, and those disease states which have been found to be usefullytreated by the specific antibacterials of this invention. Such diseasestates include, but are not limited to, treatment of a mammal afflictedwith pathogenic bacteria, in particular staphylococci (methicillinsensitive and resistant), streptococci (penicillin sensitive andresistant), enterococci (vancomycin sensitive and resistant), andClostridium difficile.

[0181] The term “therapeutically effective amount” refers to that amountwhich is sufficient to effect treatment, as defined herein, whenadministered to a mammal in need of such treatment. The therapeuticallyeffective amount will vary depending on the subject and disease statebeing treated, the severity of the affliction and the manner ofadministration, and may be determined routinely by one of ordinary skillin the art.

[0182] The term “protecting group” or “blocking group” refers to anygroup which, when bound to one or more hydroxyl, thiol, amino, carboxyor other groups of the compounds, prevents undesired reactions fromoccurring at these groups and which protecting group can be removed byconventional chemical or enzymatic steps to reestablish the hydroxyl,thio, amino, carboxy or other group. The particular removable blockinggroup employed is not critical and preferred removable hydroxyl blockinggroups include conventional substituents such as allyl, benzyl, acetyl,chloroacetyl, thiobenzyl, benzylidine, phenacyl, t-butyl-diphenylsilyland any other group that can be introduced chemically onto a hydroxylfunctionality and later selectively removed either by chemical orenzymatic methods in mild conditions compatible with the nature of theproduct. Protecting groups are disclosed in more detail in T. W. Greeneand P. G. M. Wuts, “Protective Groups in Organic Synthesis” 3^(rd) Ed.,1999, John Wiley and Sons, N.Y.

[0183] Preferred removable amino blocking groups include conventionalsubstituents such as t-butyoxycarbonyl (t-BOC), benzyloxycarbonyl (CBZ),fluorenylmethoxycarbonyl (FMOC), allyloxycarbonyl (ALOC) and the like,which can be removed by conventional conditions compatible with thenature of the product.

[0184] Preferred carboxy protecting groups include esters such asmethyl, ethyl, propyl, t-butyl etc. which can be removed by mildconditions compatible with the nature of the product.

[0185] General Synthetic Procedures

[0186] The glycopeptide compounds of this invention can be prepared fromreadily available starting materials using the following general methodsand procedures. It will be appreciated that where typical or preferredprocess conditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

[0187] Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and G. M. Wuts, ProtectingGroups in Organic Synthesis, Third Edition, Wiley, N.Y., 1999, andreferences cited therein.

[0188] In the following reaction schemes, the glycopeptide compounds aredepicted in a simplified form as a box “G” that shows the carboxyterminus labeled [C], the vancosamine amino terminus labeled [V], the“non-saccharide” amino terminus (leucine amine moiety) labeled [N], andoptionally, the resorcinol moiety labeled [R] as follows:

[0189] A glycopeptide compound of the present invention that issubstituted at the C-terminus with a substituent that comprises two ormore carboxy groups can be prepared by coupling the correspondingglycopeptide compound wherein the C-terminus is a carboxy group with therequisite carboxy-protected compound. Subsequent removal of the carboxyprotecting groups affords the compound of the invention. For example aglycopeptide compound of formula I wherein R³ is a nitrogen linkedmoiety comprising two or more carboxy groups can be prepared by couplinga corresponding glycopeptide compound of formula I wherein R³ is hydroxywith the requisite carboxy-protected amine to form an amide. Subsequentremoval of the carboxy protecting groups affords the compound of theinvention.

[0190] A glycopeptide compound of the present invention wherein theC-terminus is substituted with a substituent that comprises two or morecarboxy groups and wherein the vancosamine amino terminus (V) is alsosubstituted, can be prepared by first reductively alkylating thecorresponding glycopeptide compound wherein the vancosamine aminoterminus (V) is the free amine (NH₂) and then coupling the correspondingglycopeptide compound with the requisite poly carboxy-protected compoundat the C-terminus. Subsequent removal of the carboxy protecting groupsaffords the compound of the invention.

[0191] By way of illustration, a glycopeptide compound, such asvancomycin, can first be reductive alkylated as shown in the followingreaction:

[0192] where A represents R^(a) minus one carbon atom and R^(a), R^(b),Y, Z and x are as defined herein. This reaction is typically conductedby first contacting one equivalent of the glycopeptide, i.e.,vancomycin, with an excess, preferably from 1.1 to 1.3 equivalents, ofthe desired aldehyde in the presence of an excess, preferably about 2.0equivalents, of a tertiary amine, such as diisopropylethylamine (DIPEA)and the like. This reaction is typically conducted in an inert diluent,such as DMF or acetonitrile/water, at ambient temperature for about 0.25to 2 hours until formation of the corresponding imine and/or hemiaminalis substantially complete. The resulting imine and/or hemiaminal istypically not isolated, but is reacted in situ with a metal hydridereducing agent, such as sodium cyanoborohydride and the like, to affordthe corresponding amine. This reaction is preferably conducted bycontacting the imine and/or hemiaminal with an excess, preferably about3 equivalents, of trifluoroacetic acid, followed by about 1 to 1.2equivalents of the reducing agent at ambient temperature in methanol oracetonitrile/water. The resulting alkylated product is readily purifiedby conventional procedures, such as precipitation and/or reverse-phaseHPLC. Surprisingly, by forming the imine and/or hemiaminal in thepresence of a trialkyl amine, and then acidifying with trifluoroaceticacid before contact with the reducing agent, the selectivity for thereductive alkylation reaction is greatly improved, i.e., reductivealkylation at the amino group of the saccharide (e.g., vancosamine) isfavored over reductive alkylation at the N-terminus (e.g., the leucinylgroup) by at least 10:1, more preferably 20:1.

[0193] The above process is a significantly improvement over previousmethods for selectively alkylating an amino saccharide group of aglycopeptide antibiotic. Thus, the present invention also provides amethod for alkylating a glycopeptide that comprises a saccharide-aminecomprising:

[0194] combining an aldehyde or ketone, a suitable base, and theglycopeptide, to provide a reaction mixture;

[0195] acidifying the reaction mixture; and

[0196] combining the reaction mixture with a suitable reducing agent, toprovide a glycopeptide that is alkylated at the saccharide-amine.Preferably, the glycopeptide comprises at least one amino group otherthan the saccharide-amine.

[0197] Preferably, the reductive alkylation at the saccharide-amine isfavored over reductive alkylation at another amino group of theglycopeptide by at least about 10:1; and more preferably, by at leastabout 15:1 or about 20:1.

[0198] The reductive alkylation process of the invention is typicallycarried out in the presence of a suitable solvent or combination ofsolvents, such as, for example, a halogenated hydrocarbon (e.g.methylene chloride), a linear or branched ether (e.g. diethyl ether,tetrahydrofuran), an aromatic hydrocarbon (e.g. benzene or toluene), analcohol (methanol, ethanol, or isopropanol), dimethylsulfoxide (DMSO),N,N-dimethylformamide, acetonitrile, water,1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidone, tetramethyl urea,N,N-dimethylacetamide, diethylformamide (DMF), 1-methyl-2-pyrrolidinone,tetramethylenesulfoxide, glycerol, ethyl acetate, isopropyl acetate,N,N-dimethylpropylene urea (DMPU) or dioxane. Preferably the alkylationis carried out in acetonitrile/water, or DMF/methanol.

[0199] Preferably the reduction (i.e. treatment with the reducing agent)is carried out in the presence of a protic solvent, such as, forexample, an alcohol (e.g. methanol, ethanol, propanol, isopropanol, orbutanol), water, or the like.

[0200] The reductive alkylation process of the invention can be carriedout at any suitable temperature from the freezing point to the refluxtemperature of the reaction mixture. Preferably the reaction is carriedout at a temperature in the range of about 0° C. to about 100° C. Morepreferably at a temperature in a range of about 0° C. to about 50° C.,or in a range of about 20° C. to about 30° C.

[0201] Any suitable base can be employed in the reductive alkylationprocess of the invention. Suitable bases include tertiary amines (e.g.diisopropylethylamine, N-methylmorpholine or triethylamine) and thelike.

[0202] Any suitable acid can be used to acidify the reaction mixture.Suitable acids include carboxylic acids (e.g. acetic acid,trichloroacetic acid, citric acid, formic acid, or trifluoroaceticacid), mineral acids (e.g. hydrochloric acid, sulfuric acid, orphosphoric acid), and the like. A preferred acid is trifluoroaceticacid.

[0203] Suitable reducing agents for carrying out reductive alkylationprocess of the invention are known in the art. Any suitable reducingagent can be employed in the methods of the invention, provided it iscompatible with the functionality present in the glycopeptide. Forexample, suitable reducing agents include sodium cyanoborohydride,triacetoxyborohydride, pyridine/borane, sodium borohydride, and zincborohydride. The reduction can also be carried out in the presence of atransition metal catalyst (e.g. palladium or platinum) in the presenceof a hydrogen source (e.g. hydrogen gas or cycloheadiene). See forexample, Advanced Organic Chemistry, Fourth Edition, John Wiley & Sons,New York (1992), 899-900.

[0204] The glycopeptide derivative resulting from the reductivealkylation can then coupled with a polycarboxy-substituted amine (R³—H)to form an amide. This reaction is illustrated by the followingreaction:

[0205] where R³ is a nitrogen-linked group that comprises at least twoprotected carboxy groups. In this reaction, the glycopeptide derivativeis typically contacted with the amine in the presence of a peptidecoupling reagent, such as PyBOP and HOBT, to provide the amide. Thisreaction is typically conducted in an inert diluent, such as DMF, at atemperature ranging from about 0° C. to about 60° C. for about 1 to 24hours or until the coupling reaction is substantially complete.Subsequent deprotection using conventional procedures and reagentsaffords the compound of this invention.

[0206] If desired, the amine coupling step described above can beconducted first to provide an amide, followed by reductive alkylationand deprotection to afford the compound of the invention.

[0207] If desired, the glycopeptide compounds of this invention can alsobe prepared in a step-wise manner in which a precursor to the—R^(a)—Y—R^(b)—(Z)_(x) group is first attached the glycopeptide byreductive alkylation, followed by subsequent elaboration of the attachedprecursor using conventional reagent and procedures to form the—R^(a)—Y—R^(b)—(Z)_(x) group. Additionally, ketones may also be employedin the above-described reductive alkylation reactions to affordα-substituted amines.

[0208] Any glycopeptide having an amino group may be employed in thesereductive alkylation reactions. Such glycopeptides are well-known in theart and are either commercially available or may be isolated usingconventional procedures. Suitable glycopeptides are disclosed, by way ofexample, in U.S. Pat. Nos. 3,067,099; 3,338,786; 3,803,306; 3,928,571;3,952,095; 4,029,769; 4,051,237; 4,064,233; 4,122,168; 4,239,751;4,303,646; 4,322,343; 4,378,348; 4,497,802; 4,504,467; 4,542,018;4,547,488; 4,548,925; 4,548,974; 4,552,701; 4,558,008; 4,639,433;4,643,987; 4,661,470; 4,694,069; 4,698,327; 4,782,042; 4,914,187;4,935,238; 4,946,941; 4,994,555; 4,996,148; 5,187,082; 5,192,742;5,312,738; 5,451,570; 5,591,714; 5,721,208; 5,750,509; 5,840,684; and5,843,889. Preferably, the glycopeptide employed in the above reactionis vancomycin.

[0209] As illustrated in the following scheme, an aminoalkyl sidechainat the resorcinol moiety of a glycopeptide, such as vancomycin, can beintroduced via a Mannich reaction (in this scheme, the resorcinol moietyis shown for clarity). In this reaction, an amine (NHR^(c)R^(c)) and analdehyde (CH₂O), such as formalin (a source of formaldehyde), arereacted with the glycopeptide under basic conditions to give theglycopeptide derivative.

[0210] Compounds of the invention comprising a sulfoxide or sulfone canbe prepared from the corresponding thio compounds using conventionalreagents and procedures. Suitable reagents for oxidizing a thio compoundto a sulfoxide include, by way of example, hydrogen peroxide, peracidessuch as 3-chloroperoxybenzoic acid (MCPBA), sodium periodate, sodiumchlorite, sodium hypochlorite, calcium hypochlorite, tert-butylhypochlorite and the like. Chiral oxidizing reagents, (optically activereagents) may also be employed to provide chiral sulfoxides. Suchoptically active reagents are well-known in the art and include, forexample, the reagents described in Kagen et al., Synlett., 1990,643-650.

[0211] The aldehydes and ketones employed in the above reactivealkylation reactions are also well-known in the art and are eithercommercially available or can be prepared by conventional proceduresusing commercially available starting materials and conventionalreagents (for example see March, Advanced Organic Chemistry, FourthEdition, John Wiley & Sons, New York (1992), and references citedtherein).

[0212] The poly-carboxy-substituted compounds (e.g.poly-carboxy-substituted amines, alcohols, and thiols) employed toprepare the compounds of the invention are either commercially availableor can be prepared by conventional procedures using commerciallyavailable starting materials and reagents. For example, the aminopolyacids L-aspartic acid, D-aspartic acid, DL-aspartic acid,N-methyl-D-aspartic acid, L-glutamic acid, D-glutamic acid, D,L-glutamicacid, DL-2-methylglutamic acid, DL-2-aminoadipic acid, D-2-aminoadipicacid, L-2-aminoadipic acid, 3-aminoadipic acid, 2,6-diaminopimelic acid,L-gamma-carboxyglutamic acid, lanthionine, D-cystine, L-cystine,iminodiacetic acid, ethylenediamine-N,N′-diacetic acid, and kainic acid,as well as numerous biscarboxy-protected derivatives of L-aspartic acidand L-glutamic acid are commercially available from Sigma, St. Louis,Mo. and Aldrich Chemical Company, Milwaukee, Wis. Otherbiscarboxy-protected derivatives of L-aspartic acid and the like, suchas the bis-fluorenylmethyl esters, are readily prepared using well-knownprocedures and reagents.

[0213] Additional details and other methods for preparing the compoundsof this invention are described in the Examples below.

[0214] Pharmaceutical Compositions

[0215] This invention also includes pharmaceutical compositioncontaining the novel glycopeptide compounds of this invention.Accordingly, the glycopeptide compound, preferably in the form of apharmaceutically acceptable salt, can be formulated for oral orparenteral administration for the therapeutic or prophylactic treatmentof bacterial infections.

[0216] By way of illustration, the glycopeptide compound can be admixedwith conventional pharmaceutical carriers and excipients and used in theform of tablets, capsules, elixirs, suspensions, syrups, wafers, and thelike. Such pharmaceutical compositions will contain from about 0.1 toabout 90% by weight of the active compound, and more generally fromabout 10 to about 30%. The pharmaceutical compositions may containcommon carriers and excipients, such as corn starch or gelatin, lactose,sucrose, microcrystalline cellulose, kaolin, mannitol, dicalciumphosphate, sodium chloride, and alginic acid. Disintegrators commonlyused in the formulations of this invention include croscarmellose,microcrystalline cellulose, corn starch, sodium starch glycolate andalginic acid.

[0217] A liquid composition will generally consist of a suspension orsolution of the compound or pharmaceutically acceptable salt in asuitable liquid carrier(s), for example ethanol, glycerine, sorbitol,non-aqueous solvent such as polyethylene glycol, oils or water,optionally with a suspending agent, a solubilizing agent (such as acyclodextrin), preservative, surfactant, wetting agent, flavoring orcoloring agent. Alternatively, a liquid formulation can be prepared froma reconstitutable powder.

[0218] For example a powder containing active compound, suspendingagent, sucrose and a sweetener can be reconstituted with water to form asuspension; and a syrup can be prepared from a powder containing activeingredient, sucrose and a sweetener.

[0219] A composition in the form of a tablet can be prepared using anysuitable pharmaceutical carrier(s) routinely used for preparing solidcompositions. Examples of such carriers include magnesium stearate,starch, lactose, sucrose, microcrystalline cellulose and binders, forexample polyvinylpyrrolidone. The tablet can also be provided with acolor film coating, or color included as part of the carrier(s). Inaddition, active compound can be formulated in a controlled releasedosage form as a tablet comprising a hydrophilic or hydrophobic matrix.

[0220] A composition in the form of a capsule can be prepared usingroutine encapsulation procedures, for example by incorporation of activecompound and excipients into a hard gelatin capsule. Alternatively, asemi-solid matrix of active compound and high molecular weightpolyethylene glycol can be prepared and filled into a hard gelatincapsule; or a solution of active compound in polyethylene glycol or asuspension in edible oil, for example liquid paraffin or fractionatedcoconut oil can be prepared and filled into a soft gelatin capsule.

[0221] Tablet binders that can be included are acacia, methylcellulose,sodium carboxymethylcellulose, poly-vinylpyrrolidone (Povidone),hydroxypropyl methylcellulose, sucrose, starch and ethylcellulose.Lubricants that can be used include magnesium stearate or other metallicstearates, stearic acid, silicone fluid, talc, waxes, oils and colloidalsilica.

[0222] Flavoring agents such as peppermint, oil of wintergreen, cherryflavoring or the like can also be used. Additionally, it may bedesirable to add a coloring agent to make the dosage form moreattractive in appearance or to help identify the product.

[0223] The compounds of the invention and their pharmaceuticallyacceptable salts that are active when given parenterally can beformulated for intramuscular, intrathecal, or intravenousadministration.

[0224] A typical composition for intra-muscular or intrathecaladministration will consist of a suspension or solution of activeingredient in an oil, for example arachis oil or sesame oil. A typicalcomposition for intravenous or intrathecal administration will consistof a sterile isotonic aqueous solution containing, for example activeingredient and dextrose or sodium chloride, or a mixture of dextrose andsodium chloride. Other examples are lactated Ringer's injection,lactated Ringer's plus dextrose injection, Normosol-M and dextrose,Isolyte E, acylated Ringer's injection, and the like. Optionally, aco-solvent, for example, polyethylene glycol; a chelating agent, forexample, ethylenediamine tetracetic acid; a solubilizing agent, forexample, a cyclodextrin; and an anti-oxidant, for example, sodiummetabisulphite, may be included in the formulation. Alternatively, thesolution can be freeze dried and then reconstituted with a suitablesolvent just prior to administration.

[0225] In a preferred embodiment, the glycopeptide derivatives of thisinvention are formulated in an aqueous solution containing acyclodextrin. In another preferred embodiment the glycopeptidederivatives of this invention are formulated as a lyophilized powdercontaining a cyclodextrin or as a sterile powder containing acyclodextrin. Preferably, the cyclodextrin ishydroxypropyl-β-cyclodextrin or sulfobutyl ether β-cyclodextrin; morepreferably, the cyclodextrin is hydroxypropyl-β-cyclodextrin. Typically,in an injectable solution, the cyclodextrin will comprise about 1 to 25weight percent; preferably, about 2 to 10 weight percent; morepreferable, about 4 to 6 weight percent, of the formulation.Additionally, the weight ratio of the cyclodextrin to the glycopeptidederivative will preferably be from about 1:1 to about 10:1.

[0226] The compounds of the invention and their pharmaceuticallyacceptable salts which are active on rectal administration can beformulated as suppositories. A typical suppository formulation willgenerally consist of active ingredient with a binding and/or lubricatingagent such as a gelatin or cocoa butter or other low melting vegetableor synthetic wax or fat.

[0227] The compounds of this invention and their pharmaceuticallyacceptable salts which are active on topical administration can beformulated as transdermal compositions or transdermal delivery devices(“patches”). Such compositions include, for example, a backing, activecompound reservoir, a control membrane, liner and contact adhesive. Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds of the present invention in controlledamounts. The construction and use of transdermal patches for thedelivery of pharmaceutical agents is well known in the art. See, e.g.,U.S. Pat. No. 5,023,252, issued Jun. 11, 1991. Such patches may beconstructed for continuous, pulsatile, or on demand delivery ofpharmaceutical agents.

[0228] The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It, willbe understood, however, that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered and itsrelative activity, the age, weight, and response of the individualpatient, the severity of the patient's symptoms, and the like.

[0229] Suitable doses are in the general range of from 0.01-100mg/kg/day, preferably 0.1-50 mg/kg/day. For an average 70 kg human, thiswould amount to 0.7 mg to 7 g per day, or preferably 7 mg to 3.5 g perday. A more preferred dose for a human is about 500 mg to about 2 g perday.

[0230] Other suitable formulations for use in the present invention canbe found in Remington's Pharmaceutical Sciences, Mace PublishingCompany, Philadelphia, Pa., 17th ed. (1985).

[0231] The following formulation examples illustrate representativepharmaceutical compositions of the present invention.

Formulation Example A

[0232] This example illustrates the preparation of a representativepharmaceutical composition for oral administration of a compound of thisinvention: Ingredients Quantity per tablet, (mg) Active Compound 200Lactose, spray-dried 148 Magnesium stearate 2

[0233] The above ingredients are mixed and introduced into a hard-shellgelatin capsule.

Formulation Example B

[0234] This example illustrates the preparation of anotherrepresentative pharmaceutical composition for oral administration of acompound of this invention: Ingredients Quantity per tablet, (mg) ActiveCompound 400 Cornstarch 50 Lactose 145 Magnesium stearate 5

[0235] The above ingredients are mixed intimately and pressed intosingle scored tablets.

Formulation Example C

[0236] This example illustrates the preparation of a representativepharmaceutical composition for oral administration of a compound of thisinvention.

[0237] An oral suspension is prepared having the following composition.Ingredients Active Compound  1.0 g Fumaric acid  0.5 g Sodium chloride 2.0 g Methyl paraben  0.1 g Granulated sugar  25.5 g Sorbitol (70%solution) 12.85 g Veegum K (Vanderbilt Co.)  1.0 g Flavoring 0.035 mlColorings  0.5 mg Distilled water q.s. to 100 ml

Formulation Example D

[0238] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0239] An injectable preparation buffered to a pH of 4 is preparedhaving the following composition: Ingredients Active Compound 0.2 gSodium Acetate Buffer Solution (0.4 M) 2.0 ml HCl (1N) q.s. to pH 4Water (distilled, sterile) q.s. to 20 ml

Formulation Example E

[0240] This example illustrates the preparation of a representativepharmaceutical composition for injection of a compound of thisinvention.

[0241] A reconstituted solution is prepared by adding 20 ml of sterilewater to 1 g of the compound of this invention. Before use, the solutionis then diluted with 200 ml of an intravenous fluid that is compatiblewith the active compound. Such fluids are chosen from 5% dextrosesolution, 0.9% sodium chloride, or a mixture of 5% dextrose and 0.9%sodium chloride. Other examples are lactated Ringer's injection,lactated Ringer's plus 5% dextrose injection, Normosol-M and 5%dextrose, Isolyte E, and acylated Ringer's injection

Formulation Example F

[0242] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0243] An injectable preparation is prepared having the followingcomposition: Ingredients Active Compound 0.1-5.0 gHydroxypropyl-β-cyclodextrin   1-25 g 5% Aqueous Dextrose Solution(sterile) q.s. to 100 ml

[0244] The above ingredients are blended and the pH is adjusted to3.5±0.5 using 0.5 N HCl or 0.5 N NaOH.

Formulation Example G

[0245] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0246] A frozen solution suitable for injection is prepared having thefollowing composition: Frozen Solution Active Compound 250 mg to 1000 mgHydroxypropyl-β-cyclodextrin 250 mg to 10 g Excipients - e.g., dextrose 0-50 g Water for Injection 10-100 ml

[0247] The weight ratio of hydroxypropyl-β-cyclodextrin to the activecompound will typically be from about 1:1 to about 10:1.

[0248] Representative Procedure: Hydroxypropyl-β-cyclodextrin andexcipients, if any, are dissolved in about 80% of the water forinjection and the active compound is added and dissolved. The pH isadjusted with 1 M sodium hydroxide to 4.7±0.3 and the volume is thenadjusted to 95% of the final volume with water for injection. The pH ischecked and adjusted, if necessary, and the volume is adjusted to thefinal volume with water for injection. The formulation is then sterilefiltered through a 0.22 micron filter and placed into a sterile vialunder aseptic conditions. The vial is capped, labeled and stored frozen.

Formulation Example H

[0249] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0250] A lyophilized powder useful for preparing an injectable solutionis prepared having the following composition: Lyophilized Powder ActiveCompound 250 mg to 1000 mg Hydroxypropyl-β-cyclodextrin 250 mg to 10 gExcipients - e.g., mannitol, sucrose and/or lactose 0-50 g Bufferagent - e.g., citrate 0-500 mg

[0251] The weight ratio of hydroxypropyl-β-cyclodextrin to the activecompound will typically be from about 1:1 to about 10:1.

[0252] Representative Procedure: Hydroxypropyl-β-cyclodextrin andexcipients and/or buffering agents, if any, are dissolved in about 60%of the water for injection. The active compound is added and dissolvedand the pH is adjusted with 1 M sodium hydroxide to 4.0-5.0 and thevolume is adjusted to 95% of the final volume with water for injection.The pH is checked and adjusted, if necessary, and the volume is adjustedto the final volume with water for injection. The formulation is thensterile filtered through a 0.22 micron filter and placed into a sterilevial under aseptic conditions. The formulation is then freeze-driedusing an appropriate lyophilization cycle. The vial is capped(optionally under partial vacuum or dry nitrogen), labeled and stored atroom temperature or under refrigeration.

Formulation Example I

[0253] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0254] A sterile powder useful for preparing an injectable solution isprepared having the following composition: Sterile Powder ActiveCompound 250 mg to 1000 mg Hydroxypropyl-β-cyclodextrin 250 mg to 10 g¹Excipients optional

[0255] The weight ratio of hydroxypropyl-β-cyclodextrin to the activecompound will typically be from about 1:1 to about 10:1.

[0256] Representative Procedure: Hydroxypropyl-β-cyclodextrin and theactive compound (and any excipients) are dispersed into an appropriatesterile container and the container is sealed (optionally under partialvacuum or dry nitrogen), labeled and stored at room temperature or underrefrigeration.

[0257] Administration of Representative Formulations H and I to aPatient

[0258] The pharmaceutical formulations described in formulation examplesH and I above can be administered intravenously to a patient by theappropriate medical personnel to treat or prevent gram-positiveinfections. For administration, the above formulations can bereconstituted and/or diluted with a diluent, such as 5% dextrose orsterile saline, as follows:

[0259] Representative Procedure: The lyophilized powder of formulationexample H (e.g., containing 1000 mg of active compound) is reconstitutedwith 20 ml of sterile water and the resulting solution is furtherdiluted with 80 ml of sterile saline in a 100 ml infusion bag. Thediluted solution is then administered to the patient intravenously over30 to 120 minutes.

Formulation Example J

[0260] This example illustrates the preparation of a representativepharmaceutical composition for topical application of a compound of thisinvention. Ingredients grams Active compound 0.2-10 Span 60 2 Tween 60 2Mineral oil 5 Petrolatum 10 Methyl paraben 0.15 Propyl paraben 0.05 BHA(butylated hydroxy anisole) 0.01 Water q.s. to 100

[0261] All of the above ingredients, except water, are combined andheated to 60° C. with stirring. A sufficient quantity of water at 60° C.is then added with vigorous stirring to emulsify the ingredients, andwater then added q.s. 100 g.

Formulation Example K

[0262] This example illustrates the preparation of a representativepharmaceutical composition containing a compound of this invention.

[0263] A suppository totaling 2.5 grams is prepared having the followingcomposition: Ingredients Active Compound 500 mg Witepsol H-15* balance

[0264] Utility

[0265] The glycopeptide compounds of this invention, and theirpharmaceutically acceptable salts, are useful in medical treatments andexhibit biological activity, including antibacterial activity, which canbe demonstrated in using the tests described herein. Such tests are wellknown to those skilled in the art, and are referenced and described inLorian “Antibiotics in Laboratory Medicine”, Fourth Edition, Williamsand Wilkins (1991).

[0266] Accordingly, this invention provides methods for treatingbacterial or infectious diseases, especially those caused byGram-positive microorganisms, in animals. The compounds of thisinvention are particularly useful in treating infections caused bymethicillin-resistant staphylococci. Also, the compounds are useful intreating infection due to enterococci, including vancomycin-resistantenterococci (VRE). Examples of such diseases include severestaphylococcal infections, such as staphylococcal endocarditis andstaphylococcal septicemia. The animal treated may be either susceptibleto, or infected with, the microorganism. The method of treatmenttypically comprises administering to the animal an amount of a compoundof this invention which is effective for this purpose.

[0267] In practicing this method, the antibiotic can be administered ina single daily dose or in multiple doses per day. The treatment regimenmay require administration over extended periods of time, for example,for several days or for from one to six weeks. The amount peradministered dose or the total amount administered will depend on suchfactors as the nature and severity of the infection, the age and generalhealth of the patient, the tolerance of the patient to the antibioticand the microorganism or microorganisms in the infection.

[0268] The following synthetic and biological examples are offered toillustrate this invention and are not to be construed in any way aslimiting the scope of this invention.

EXAMPLES

[0269] In the examples below, the following abbreviations have thefollowing meanings. Any abbreviations not defined have their generallyaccepted meaning. Unless otherwise stated, all temperatures are indegrees Celsius.

[0270] ACN=acetonitrile

[0271] BOC, Boc=tert-butoxycarbonyl

[0272] DIBAL-H=diisobutylaluminum hydride

[0273] DIPEA=diisopropylethylamine

[0274] DMF=N,N-dimethylformamide

[0275] DMSO=dimethyl sulfoxide

[0276] eq.=equivalent

[0277] EtOAc=ethyl acetate

[0278] Fmoc=9-fluorenylmethoxycarbonyl

[0279] HOBT=1-hydroxybenzotriazole hydrate

[0280] Me=methyl

[0281] PyBOP=benzotriazol-1-yloxytris(pyrrolidino)phosphoniumhexafluorophosphate

[0282] TEMPO=2,2,6,6-tetramethyl-piperidinyloxy, free radical

[0283] TFA=trifluoroacetic acid

[0284] THF=tetrahydrofuran

[0285] TLC, tlc=thin layer chromatography

[0286] In the following examples, vancomycin hydrochloride semi-hydratewas purchased from Alpharma, Inc. Fort Lee, N.J. 07024 (Alpharma AS,Oslo Norway). Other reagents and reactants are available from AldrichChemical Co., Milwaukee, Wis. 53201.

[0287] General Procedure A

[0288] Reductive Alkylation of Vancomycin

[0289] To a mixture of vancomycin (1 eq.) and the desired aldehyde (1.3eq.) in DMF was added DIPEA (2 eq.). The reaction was stirred at ambienttemperature for 1-2 hours and monitored by reverse-phase HPLC. Methanoland NaCNBH₃ (1 eq.) were added to the solution, followed by TFA (3 eq.).Stirring was continued for an additional hour at ambient temperature.After the reaction was complete, the methanol was removed in vacuo. Theresidue was precipitated in acetonitrile. Filtration gave the crudeproduct which was then purified by reverse-phase HPLC. If desired, otherglycopeptides antibiotics may be used in this procedure.

[0290] General Procedure B

[0291] Synthesis of S-Decyl Mercaptoacetaldehyde

[0292] Under nitrogen, to a suspension of potassium carbonate (27 g, 200mmol) in acetone (100 ml) was added decyl bromide (10 ml, 50 mmol) andmercaptoethanol (4.4 ml, 63 mmol). The suspension was stirred at roomtemperature for 2 days, then partitioned between water and 80%hexane/ethyl acetate. The organic phase was washed with 2N sodiumhydroxide, dried over magnesium sulfate, and the volatiles removed undervacuum to give S-decyl mercaptoethanol (10.2 g, 47 mmol) as a/colorlessliquid that was used without further purification.

[0293] Under nitrogen, S-decyl mercaptoethanol (50 g, 230 mmol),N,N-diisopropylethylamine (128 ml, 730 mmol) and methylene chloride (400ml) were cooled to −40° C. To this solution was added, over 15 minutes,a solution of sulfur trioxide pyridine complex (116 g, 730 mmol) indimethyl sulfoxide (600 ml) and methylene chloride (200 ml). Afteraddition, the mixture was stirred a further 15 minutes at −40° C., then600 ml ice water as added. The mixture was removed from the coolingbath, 1 L water was added, and the liquids partitioned. The organicphase was washed with 1 L of 1 N hydrochloric acid, and dried overmagnesium sulfate. Filtration gave 600 ml liquid, which was diluted with600 ml hexane and passed through 200 ml silica. The silica was washedwith 100 ml 50% methylene chloride/hexane, then 300 ml methylenechloride. The combined organics were concentrated in vacuo to giveS-decyl mercaptoacetaldehyde (48 g, 220 mmol) as a colorless liquid thatwas used without further purification.

Example 1

[0294] Preparation of Compound 5

[0295] (Formula II wherein R¹⁷ is —CH(COOH)CH₂COOH, R¹⁸ is hydrogen, R¹⁹is hydrogen, and R²⁰ is —CH₂CH₂—S—(CH₂)₉CH₃)

[0296] Under nitrogen, to a solution of S-decyl mercaptoacetaldehyde(crude, 48 g, 220 mmol) in N,N-dimethylformamide (1.4 L) was added solidvancomycin hydrochloride hydrate (173 g, 110 mmol) followed byN,N-diisopropylethylamine (58 ml, 330 mmol). The suspension was stirredvigorously at room temperature for 2 hours, in the course of which timeall the vancomycin fully dissolved. Then trifluoroacetic acid (53 ml,690 mmol) was added. The solution was stirred a further 90 minutes. Thensolid sodium cyanoborohydride (10.5 g, 170 mmol) followed by methanol(800 mL) were added. After three hours, the mixture was poured intowater (7 L), resulting in a slightly cloudy solution. The pH of thesolution was adjusted to 5 with saturated sodium bicarbonate, resultingin the formation of a white precipitate. This precipitate was collectedby filtration, washed with water, then ethyl acetate, and dried undervacuum to give a compound of formula II wherein R¹⁷ is OH; R¹⁸ ishydrogen, R¹⁹ is hydrogen, and R²⁰ is —CH₂CH₂—S—(CH₂)₉CH₃) (compound a).

[0297] A solution of benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate (PyBOP, 16.5 g, 32 mmol) in N,N-dimethylformamide(DMF, 82.5 ml) was prepared. Under nitrogen, compound a from above,(crude, 57 g) was dissolved in DMF (250 ml) at room temperature.1-Hydroxybenzotriazole (4.7 g, 35 mmol) was added, then L-aspartic acidbis-fluorenylmethyl ester trifluoroacetate (17 g, 28 mmol), followed byN,N-diisopropylethylamine (18 ml, 105 mmol). To this solution was added54 ml of the PyBOP solution, and the mixture stirred for 35 minutes.Then a further 16.5 ml of the PyBOP solution was added. After another 30minutes, 8.6 ml more PyBOP solution was added. 35 minutes later, thefinal 1.1 ml of the PyBOP solution was added. The progress of thereaction over this time was monitored by reverse-phase HPLC. 55 minutesafter the final PyBOP addition, piperidine (37 ml, 375 mmol) was addedand the solution stirred for 25 minutes at room temperature. The mixturewas then concentrated under vacuum to around 100 ml volume, then addedto 2.5 L acetonitrile. The resulting precipitate was stirred overnight,filtered, washed with acetonitrile, and dried under vacuum to give thetitle compound, which was purified by reverse-phase HPLC.

Example 2

[0298] Preparation of Compound 7

[0299] (Formula II wherein R¹⁷ is —CH(COOH)CH₂COOH, R¹⁸ is hydrogen, R¹⁹is hydrogen, and R²⁰ is —CH₂CH₂O(CH₂)₉CH₃

[0300] N^(VAN)-decyloxoethyl Vancomycin bistrifluoroacetate (3 g, 1.6mmol) and NH₂-Asp (OFm)-(OFm) trifluoroacetate (1.5 g, 2.4 mmol) wasdissolved in anhydrous DMF (30 ml). To this solution was added DIPEA(1.5 ml, 8.9 mmol). After 5 minutes stirring, a solution of PyBOP (0.94g, 1.8 mmol) and HOBT (0.28 g, 1.8 mmol) in anhydrous DMF (1 ml) wasadded and the reaction was stirred for 1.5 hours. The solution was thenpoured into diethyl ether (150 ml) and the resulting solid was filtered,washed with diethyl ether (3×30 ml) and dried under reduced pressure.Once dry, the white solid was re-dissolved in DMF (25 ml) and piperidine(5.5 ml) and stirred for 45 minutes. The solution was then poured intodiethyl ether (150 ml) and the resulting solid was filtered, washed withMeCN (2×50 ml), diethyl ether (2×50 ml), and dried under reducedpressure. The crude product was purified by reverse-phase HPLC to givethe title compound. MS calculated (M+) 1748.7; found (MH+) 1749.7.

Example 3

[0301] Preparation of Compound 15

[0302] (Formula II wherein NR¹⁷R¹⁸ is formula III wherein R^(g) isN-(D-glucamine), R¹⁹ is hydrogen, and R²⁰ is —CH₂CH₂O(CH₂)₉CH₃

[0303] To a mixture of N^(VAN)-decyloxoethyl Vancomycinbistrifluoroacetate (2 g, 1.1 mmol) in DMF (20 ml) was added a solutionof PyBOP (0.61 g, 1.2 mmol) and HOBT (0.16 g, 1.2 mmol) in DMF (2 ml)followed by NMM (0.12 ml, 1.1 mmol). After 30 minutes, additional NMMwas added (0.13 ml, 1.2 mmol). After 10 minutes a solution of NH₂-Glu(NH-Me-D-Glucamine)-OH hydrochloride and NMM (0.35 ml, 3.3 mmol) in DMF(4 ml) was added and the reaction was stirred for 2 hours. The thickreaction was then diluted with DMF (10 ml) and NH₂-Asp(OFm)-O(Fm)trifluoroacetate (3.2 g, 5.5 mmol), DIPEA (1.1 ml, 6.6 mmol), and asolution of PyBOP (0.61 g, 1.2 mmol) and HOBT (0.16 g, 1.2 mmol) in DMF(2 ml) were added sequentially. After 1 hour of stirring at roomtemperature additional PyBOP (0.31 g, 0.6 mmol) and HOBT (0.09 g, 0.6mmol) in DMF (2 ml) was added. The reaction was stirred for 20 minutesand then added to diethyl ether (250 ml). The resulting precipitate wasfiltered, washed with MeCN (50 ml), diethyl ether (50 ml), and driedunder reduced pressure to give 3.1 g white solid.

[0304] The crude solid was taken up in DMF (15 ml) and piperidine (4 ml)and stirred for 30 minutes. The reaction was then added to diethyl etherand the resulting solid was filtered, dried under reduced pressure, andthen purified by reverse phase HPLC to give the titled compound. MScalculated (M+) 2055.0; found (MH+) 2055.9.

[0305] Using the above procedures and the appropriate starting materialsthe compounds shown in Table I were prepared. The mass spectral data forthese compounds were as follows: Compound No. MW (freebase) Observed MH*1 1704.6 1705.9 2 1718.7 1719.4 3 1732.7 1733.2 4 1746.7 1747.9 5 1764.81765.6 6 1761.7 7 1748.7 1749.7 8 1765.0 1765.6 9 1825.1 1825.7 101809.1 1809.7 11 1807.11 1807.8 12 1762.7 1764.1 13 2055.0 14 2054.02054.7 15 2055 2055.9

Example 4

[0306] Determination of Antibacterial Activity

[0307] A. In vitro Determination of Antibacterial Activity

[0308] 1. Determination of Minimal Inhibitory Concentrations (MICs)

[0309] Bacterial strains were obtained from either American Type TissueCulture Collection (ATCC), Stanford University Hospital (SU), KaiserPermanente Regional Laboratory in Berkeley (KPB), Massachusetts GeneralHospital (MGH), the Centers for Disease Control (CDC), the San FranciscoVeterans' Administration Hospital (SFVA) or the University of CaliforniaSan Francisco Hospital (UCSF). Vancomycin resistant enterococci werephenotyped as Van A or Van B based on their sensitivity to teicoplanin.Some vancomycin resistant enterococci that had been genotyped as Van A,Van B, Van C1 or Van C2 were obtained from the Mayo Clinic.

[0310] Minimal inhibitory concentrations (MICS) were measured in amicrodilution broth procedure under NCCLS guidelines. Routinely, thecompounds were serially diluted into Mueller-Hinton broth in 96-wellmicrotiter plates. Overnight cultures of bacterial strains were dilutedbased on absorbance at 600 nm so that the final concentration in eachwell was 5×10⁵ cfu/ml. Plates were returned to a 35° C. incubator. Thefollowing day (or 24 hours in the case of Enterococci strains), MICswere determined by visual inspection of the plates. Strains routinelytested in the initial screen included methicillin-sensitiveStaphylococcus aureus (MSSA), methicillin-resistant Staphylococcusaureus, methicillin-sensitive Staphylococcus epidermidis (MSSE),methicillin-resistant Staphylococcus epidermidis (MRSE), vancomycinsensitive Enterococcus faecium (VSE Fm), vancomycin sensitiveEnterococcus faecalis (VSE Fs), vancomycin resistant Enterococcusfaecium also resistant to teicoplanin (VRE Fm Van A), vancomycinresistant Enterococcus faecium sensistive to teicoplanin (VRE Fm Van B),vancomycin resistant Enterococcus faecalis also resistant to teicoplanin(VRE Fs Van A), vancomycin resistant Enterococcus faecalis sensitive toteicoplanin (VRE Fs Van B), enterococcus gallinarium of the Van Agenotype (VRE Gm Van A), enterococcus gallinarium of the Van C-1genotype (VRE Gm Van C-1), enterococcus casseliflavus of the Van C-2genotype (VRE Cs Van C-2), enterococcus flavescens of the Van C-2genotype (VRE Fv Van C-2), and penicillin-sensitive Streptococcuspneumoniae (PSSP) and penicillin-resistant Streptococcus pneumoniae(PSRP). Because of the inability of PSSP and PSRP to grow well inMueller-Hinton broth, MICs with those strains were determined usingeither TSA broth supplemented with defibrinated blood or blood agarplates. Compounds which had significant activity against the strainsmentioned above were then tested for MIC values in a larger panel ofclinical isolates including the species listed above as well asnon-speciated coagulase negative Staphylococcus both sensitive andresistant to methicillin (MS-CNS and MR-CNS). In addition, they weretested for MICs against gram negative organisms, such as Escherichiacoli and Pseudomonas aeruginosa.

[0311] 2. Determination of Kill Time

[0312] Experiments to determine the time required to kill the bacteriawere conducted as described in Lorian, “Antibiotics in LaboratoryMedicine”, Fourth Edition, Williams and Wilkins (1991). Theseexperiments were conducted normally with both staphylococcus andenterococcus strains.

[0313] Briefly, several colonies were selected from an agar plate andgrown at 35° C. under constant agitation until it achieved a turbidityof approximately 1.5 and 10⁸ CFU/ml. The sample was then diluted toabout 6×10⁶ CFU/ml and incubated at 35° C. under constant agitation wascontinued. At various times aliquots were removed and five ten-foldserial dilutions were performed. The pour plate method was used todetermine the number of colony forming units (CFUs).

[0314] In general, the compounds of the invention were active in theabove tests in vitro tests and demonstrated a broad spectrum ofactivity.

[0315] B. In vivo Determination of Antibacterial Activity

[0316] 1. Acute Tolerability Studies in Mice

[0317] In these studies, a compound of this invention was administeredeither intravenously or subcutaneously and observed for 5-15 minutes. Ifthere were no adverse effects, the dose was increased in a second groupof mice. This dose incrementation continued until mortality occurred, orthe dose was maximized. Generally, dosing began at 20 mg/kg andincreased by 20 mg/kg each time until the maximum tolerated dose (MTD)is achieved.

[0318] 2. Bioavailability Studies in Mice

[0319] Mice were administered a compound of this invention eitherintravenously or subcutaneously at a therapeutic dose (in general,approximately 50 mg/kg). Groups of animals were placed in metaboliccages so that urine and feces could be collected for analysis. Groups ofanimals (n=3) were sacrificed at various times (10 min, 1 hour and 4hours). Blood was collected by cardiac puncture and the following organswere harvested—lung, liver, heart, brain, kidney, and spleen. Tissueswere weighed and prepared for HPLC analysis. HPLC analysis on the tissuehomogenates and fluids was used to determine the concentration of thetest compound or IiI present. Metabolic products resulting from changesto the test compound were also determined at this juncture.

[0320] 3. Mouse Septicemia Model

[0321] In this model, an appropriately virulent strain of bacteria (mostcommonly S. aureus, or E. Faecalis or E. Faecium) was administered tomice (N=5 to 10 mice per group) intraperitoneally. The bacteria wascombined with hog gastric mucin to enhance virulence. The dose ofbacteria (normally 10⁵-10⁷) was that sufficient to induce mortality inall of the mice over a three day period. One hour after the bacteria wasadministered, a compound of this invention was administered in a singledose either IV or subcutaneously. Each dose was administered to groupsof 5 to 10 mice, at doses that typically ranged from a maximum of about20 mg/kg to a minimum of less than 1 mg/kg. A positive control (normallyvancomycin with vancomycin sensitive strains) was administered in eachexperiment. The dose at which approximately 50% of the animals are savedwas calculated from the results.

[0322] 4. Neutropenic Thigh Model

[0323] In this model, antibacterial activity of a compound of thisinvention was evaluated against an appropriately virulent strain ofbacteria (most commonly S. aureus, or E. Faecalis or E. Faecium,sensitive or resistant to vancomycin). Mice were initially renderedneutropenic by administration of cyclophosphamide at 200 mg/kg on day 0and day 2. On day 4 they were infected in the left anterior thigh by anIM injection of a single dose of bacteria. The mice were thenadministered the test compound one hour after the bacteria and atvarious later times (normally 1, 2.5, 4 and 24 hours) the mice weresacrificed (3 per time point) and the thigh excised, homogenized and thenumber of CFUs (colony forming units) were determined by plating. Bloodwas also plated to determine the CFUs in the blood.

[0324] 5. Pharmacokinetic Studies

[0325] The rate at which a compound of this invention is removed fromthe blood can be determined in either rats or mice. In rats, the testanimals were cannulated in the jugular vein. The test compound wasadministered via tail vein injection, and at various time points(normally 5, 15, 30, 60 minutes and 2, 4, 6 and 24 hours) blood waswithdrawn from the cannula In mice, the test compound was alsoadministered via tail vein injection, and at various time points. Bloodwas normally obtained by cardiac puncture. The concentration of theremaining test compound was determined by HPLC.

[0326] In general, the compounds of the invention were active in theabove test in vivo and demonstrated a broad spectrum of activity.

Example 5

[0327] Determination of Tissue Accumulation

[0328] A. Tissue Distribution Using Radiolabeled Compound

[0329] This procedure is used to examine the tissue distribution,excretion and metabolism of a radiolabeled test compound in both maleand female rats following intravenous infusion at 10 mg/kg. Male andfemale Sprague-Dawley rats (n=2 per sex per compound) are dosed with³H-labeled test compound at 10 (400 μCi/kg) and 12.5 mg/kg (100 μCi/kg),respectively, via intravenous infusion (˜2 min). The test compound isformulated in 5% hydroxypropyl-β-cyclodextrin as 2.5 mg/ml solution.Urine and feces are cage collected over 24 hours period. At 24 hoursafter dosing, animals are sacrificed and tissues are removed. Serum,urine and tissues are analyzed for total radioactivity by oxidationfollowed by liquid scintillation counting. Urine and selected tissuessamples are extracted and analyzed by reverse phase HPLC withradioactive flow detector for the presence of potential metabolites.

[0330] B. Tissue Accumulation Following Single Dose

[0331] This procedure is used to evaluate tissue distribution of a testcompound in rats following single dose administration by infusion. MaleSprague-Dawley rats (n=3 per dose groups) are dosed with 50 mg/kg of atest compound. Two formulations are used: 30% PEG 400 and 10%sulfobutylether-β-cyclodextrin. Urine samples are cage collected over 24hours. Blood samples are collected for serum chemistry and concentrationdetermination. Liver and kidneys are removed for histology evaluation.One kidney and part of the liver are homogenized for concentrationanalysis using reverse phase HPLC with UV detection. Drug concentrationsin urine and serum samples are determined by LC-MS analysis.

[0332] C. Tissue Distribution Following Multiple Doses

[0333] This procedure is used too evaluate the potential tissueaccumulation of a test compound in rats following multiple doseadministration by intravenous infusion. Male and female Sprague-Dawleyrats (n=4 per sex per dose group) are dosed with a test compound at12.5, 25 and 50 mg/kg per day for seven days. Animals are sacrificed atday 1 (n=3 per sex per dose group) following the last dose administered.One animal per sex per dose group is retained as recovery animal andsacrificed at day 7 following the last dose administered. The testcompound is formulated in 5% hydroxypropyl-β-cyclodextrin or 1%sucrose/4.5% dextrose. Urine samples are cage collected at days 1 and 7post-dose. Blood samples are collected for serum chemistry andconcentration determination. Liver and kidneys are removed for histologyevaluation. One kidney and part of the liver are homogenized forconcentration analysis using reverse phase HPLC with UV detection. Drugconcentrations in urine and serum samples are determined by LC-MSanalysis.

[0334] While the present invention has been described with reference tothe specific embodiments thereof, it should be understood by thoseskilled in the art that various changes may be made and equivalents maybe substituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto. Additionally, all publications, patents, andpatent documents cited hereinabove are incorporated by reference hereinin full, as though individually incorporated by reference.

What is claimed is:
 1. A glycopeptide substituted at the C-terminus witha substituent that comprises two or more carboxy groups; or apharmaceutically acceptable salt, or stereoisomer, or prodrug thereof;provided the glycopeptide is not 1) teicoplanin A2 substituted at theC-terminus with a nitrogen-linked glutamic acid, 2) teicoplanin aglycon(TD) substituted at the C-terminus with a nitrogen-linked glutamic acid;or 3) a compound of formula II:

a) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ is hydrogen; R¹⁹is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ is hydrogen; b) whereinNR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ is hydrogen; R¹⁹ is hydrogen;R²⁰ is 2-(9-hydroxydecylamino)ethyl; and R²¹ is hydrogen; c) wherein R¹⁷is 1,4-dicarboxybutyl; R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is2-(decylamino)ethyl; and R²¹ is hydrogen; d) wherein NR¹⁷ isnitrogen-linked aspartic acid; R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is2-(decylamino)ethyl; and R²¹ is —CH₂—N-(D-glucamine); e) wherein R¹⁷ isnitrogen-linked aspartic acid; R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is2-[4-(4-chlorobenzyloxy)benzylamino]ethyl; and R²¹ is hydrogen; f)wherein NR¹⁷ is5-(2-carboxypyrrolidin-1-ylcarbonyl)-5-(2-carboxy-3-phenylpropylamino)pentylamino;R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ ishydrogen; g) wherein NR¹⁷ is nitrogen-linked aspartic acid; R¹⁸ ishydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; and R²¹ is—CH₂—N—(N—CH₃-D-glucamine); h) wherein NR¹⁷ is nitrogen-linked asparticacid; R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is 2-(decylamino)ethyl; andR²¹ is N-[2-(2-hydroxyethoxy)ethyl]-aminomethyl; or i) wherein NR¹⁷ isnitrogen-linked aspartic acid; R¹⁸ is hydrogen; R¹⁹ is hydrogen; R²⁰ is2-(4-isobutylbenzyl)ethyl; and R²¹ isN-[2-(2-hydroxyethoxy)ethyl]aminomethyl.
 2. The glycopeptide of claim 1wherein the substituent comprises two carboxy groups.
 3. Theglycopeptide of claim 2 wherein the substituent is a nitrogen-linkedaspartic acid or a nitrogen linked glutamic acid.
 4. The glycopeptide ofclaim 1 which is a compound of formula I:

wherein: R¹ is selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl, heterocyclic and —R^(a)—Y—R^(b)—(Z)_(x);or R¹ is a saccharide group optionally substituted with—R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); R² is hydrogen or a saccharide groupoptionally substituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f),or —C(O)—R^(a)—Y—R^(b)—(Z)_(x); R³ is a nitrogen-linked, oxygen-linked,or sulfur-linked substituent comprising two or more carboxy groups; R⁴is selected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), —C(O)R^(d) and a saccharide group optionallysubstituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); R⁵ is selected from the group consisting ofhydrogen, halo, —CH(R^(c))—NR^(c)R^(c), —CH(R^(c))—NR^(c)R^(c),—CH(R^(c))—R^(x), —CH(R^(c))—NR^(c)—Ra—C(═O)—R^(x), and—CH(R)^(c)—NR^(c)R^(a)—Y—R^(b)—(Z)_(x); R⁶ is selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, —R^(a)—Y—R^(b)—(Z)_(x),—C(O)R^(d) and a saccharide group optionally substituted with—NR^(c)—R^(a)—Y—R^(b)—(Z)_(x), or R⁵ and R⁶ can be joined, together withthe atoms to which they are attached, form a heterocyclic ringoptionally substituted with —NR^(c)—R^(a)—Y—R^(b)—(Z)_(x); R⁷ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,—R^(a)—Y—R^(b)—(Z)_(x), and —C(O)R^(d); R⁸ is selected from the groupconsisting of hydrogen, alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl andheterocyclic; R⁹ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic; R¹⁰ isselected from the group consisting of hydrogen, alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, heteroaryl and heterocyclic; or R⁸ and R¹⁰ arejoined to form —Ar¹—O—Ar²—, where Ar¹ and Ar² are independently aryleneor heteroarylene; R¹¹ is selected from the group consisting of hydrogen,alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, heteroaryl and heterocyclic, or R¹⁰ andR¹¹ are joined, together with the carbon and nitrogen atoms to whichthey are attached, to form a heterocyclic ring; R¹² is selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, heterocyclic, —C(O)R^(d), —C(NH)R^(d), —C(O)NR^(c)R^(c),—C(O)OR^(d), —C(NH)NR^(c)R^(c) and —R^(a)—Y—R^(b)—(Z)_(x), or R¹¹ andR¹² are joined, together with the nitrogen atom to which they areattached, to form a heterocyclic ring; R¹³ is selected from the groupconsisting of hydrogen or —OR¹⁴; R¹⁴ is selected from hydrogen,—C(O)R^(d) and a saccharide group; each R^(a) is independently selectedfrom the group consisting of alkylene, substituted alkylene, alkenylene,substituted alkenylene, alkynylene and substituted alkynylene; eachR^(b) is independently selected from the group consisting of a covalentbond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen; each R^(c) is independently selectedfrom the group consisting of hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl, heterocyclic and —C(O)R^(d); each R^(d) is independentlyselected from the group consisting of alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,heteroaryl and heterocyclic; R^(e) is a saccharide group; each R^(f) isindependently alkyl, substituted alkyl, alkenyl, substituted alkenyl,alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, orheterocyclic; R^(x) is an N-linked amino saccharide or an N-linkedheterocyclic; X¹, X² and X³ are each independently selected fromhydrogen or chloro; each Y is independently selected from the groupconsisting of oxygen, sulfur, —S—S—, —NR^(c)—, —S(O)—, —SO₂—,—NR^(c)C(O)—, —OSO₂—, —OC(O)—, —NR^(c)SO₂—, —C(O)NR^(c)—, —C(O)O—,—SO₂NR^(c)—, —SO₂O—, —P(O)(OR^(c))—, —P(O)(OR^(c))NR^(c)—,—OP(O)(OR^(c))O—, —OP(O)(OR^(c))NR^(c)—, —OC(O)O—, —NR^(c)C(O)O—,—NR^(c)C(O)NR^(c)—, —OC(O)NR^(c)—, —C(═O)—, and —NR^(c)SO₂NR^(c)—; eachZ is independently selected from hydrogen, aryl, cycloalkyl,cycloalkenyl, heteroaryl and heterocyclic; n is 0, 1 or 2; and x is 1 or2; or a pharmaceutically acceptable salt, or stereoisomer, or prodrugthereof.
 5. The glycopeptide of claim 4 wherein R¹ is a saccharide groupoptionally substituted with —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f),or —C(O)R^(a)—Y—R^(b)—(Z).
 6. The glycopeptide of claim 4 wherein R¹ isa saccharide group of the formula:

wherein R¹⁵ is —R^(a)—Y—R^(b)—(Z)_(x), R^(f), —C(O)R^(f), or—C(O)—R^(a)—Y—R^(b)—(Z)_(x); and R¹⁶ is hydrogen or methyl.
 7. Theglycopeptide of claim 6 wherein R¹⁵ is —CH₂CH₂—NH—(CH₂)₉CH₃;—CH₂CH₂CH₂—NH—(CH₂)₈CH₃; —CH₂CH₂CH₂CH₂—NH—(CH₂)₇CH₃;—CH₂CH₂—NHSO₂—(CH₂)₉CH₃; —CH₂CH₂—NHSO₂—(CH₂)₁₁CH₃; —CH₂CH₂—S—(CH₂)₈CH₃;—CH₂CH₂—S—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₁₀CH₃; —CH₂CH₂CH₂—S—(CH₂)₈CH₃;—CH₂CH₂CH₂—S—(CH₂)₉CH₃; —CH₂CH₂CH₂—S—(CH₂)₃—CH═CH—(CH₂)₄CH₃(trans);—CH₂CH₂CH₂CH₂—S—(CH₂)₇CH₃; —CH₂CH₂—S(O)—(CH₂)₉CH₃; —CH₂CH₂—S—(CH₂)₆Ph;—CH₂CH₂—S—(CH₂)₈Ph; —CH₂CH₂CH₂—S—(CH₂)₈Ph;—CH₂CH₂—NH—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂—NH—CH₂-4-[4-(CH₃)₂CHCH₂—]-Ph;—CH₂CH₂—NH—CH₂-4-(4-CF₃-Ph)-Ph; —CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph; —CH₂CH₂CH₂—S—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S(O)—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—S—CH₂-4-[3,4-di-Cl-PhCH₂O—)-Ph;—CH₂CH₂—NHSO₂—CH₂-4-[4-(4-Ph)-Ph]-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(4-Cl-Ph)-Ph;—CH₂CH₂CH₂—NHSO₂—CH₂-4-(Ph-C≡C—)-Ph; —CH₂CH₂CH₂—NHSO₂-4-(4-Cl-Ph)-Ph; or—CH₂CH₂CH₂—NHSO₂-4-(naphth-2-yl)-Ph.
 8. The glycopeptide of claim 6wherein R³ comprises two carboxy groups.
 9. The glycopeptide of claim 8wherein R³ is a nitrogen-linked aspartic acid or a nitrogen linkedglutamic acid.
 10. The glycopeptide of claim 6 wherein R³ is anitrogen-linked radical of formula III:

wherein R^(g) is a saccharide group.
 11. The glycopeptide of claim 10wherein R^(g) is N-(D-glucamine) or N-(D-glucosamine).
 12. Theglycopeptide of claim 4 which is a compound of formula II:

wherein: R¹⁷ is a dicarboxy-substituted alkyl group having from 3 to 10carbon atoms; R¹⁸ is selected from the group consisting of hydrogen andalkyl; R¹⁹ is hydrogen; R²⁰ is —R^(a)—Y—R^(b)—(Z)_(x); R²¹ is hydrogenR^(a) is selected from the group consisting of alkylene, substitutedalkylene, alkenylene, substituted alkenylene, alkynylene and substitutedalkynylene; R^(b) is selected from the group consisting of a covalentbond, alkylene, substituted alkylene, alkenylene, substitutedalkenylene, alkynylene and substituted alkynylene, provided R^(b) is nota covalent bond when Z is hydrogen; Y is selected from the groupconsisting of sulfur, —S(O)— and —SO₂—; each Z is independently selectedfrom hydrogen, aryl, cycloalkyl, cycloalkenyl, heteroaryl andheterocyclic; and x is 1 or 2; or a pharmaceutically acceptable salt, orstereoisomer, or prodrug thereof.
 13. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier and a therapeuticallyeffective amount of a compound of claim
 1. 14. The pharmaceuticalcomposition of claim 13, which comprises a cyclodextrin.
 15. A method oftreating a mammal having a bacterial disease, the method comprisingadministering to the mammal a therapeutically effective amount of aglycopeptide of claim
 1. 16. A method of treating a mammal having abacterial disease, the method comprising administering to the mammal atherapeutically effective amount of a glycopeptide of claim
 4. 17. Amethod of treating a mammal having a bacterial disease, the methodcomprising administering to the mammal a therapeutically effectiveamount of a glycopeptide of claim
 12. 18. A method of treating a mammalhaving a bacterial disease, the method comprising administering to themammal a therapeutically effective amount of a pharmaceuticalcomposition of claim 13.